PowerPoint 演示文稿

第一节<= span lang=3DZH-CN style=3D'color:red;mso-ansi-language:EN-US'> 基因诊断

n一、&= #22522;因诊断的概念和特= 857;

n二、&= #22522;因诊断的常用技术= 041;法

n三、&= #22522;因诊断的应用

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（一）基因诊断的= 010;念: 利用现代分子生物学技术= 644;分子遗传学= 的技术方法，直接= 816;测基因结构= 及其表达水平是否= 491;常，从而对= 疾病作出诊断的方= 861;。

n一、基因#= 786;断的概念和特点

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（二）基= ;因诊断的特点
= 3;

n针对性强= ;

n特异性高= ;

n灵敏度高= ;

n适用性强= ;

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二、基因#= 786;断的常用技术方法

n（一）、核= ;酸分子杂交技术

n（二）、聚ࡧ= 2;酶链反应(PCR)

n（三）、基ࢰ= 0;测序

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（一）核酸࠳= 8;子杂交技术

n限制性= 内切酶酶谱分析法

nDNA限制性片段长度多态= ;性分析 =

n等位基= 因特异寡核苷酸探针杂交法

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=

1、限制性内切酶酶&#= 35889;分析法

基本原理:= 利用内切= ;酶和特异性DNA= 探针来检= ;测是否存在致病基因&#= 13;

结构基因突变(点突变、= ;缺失与插= 入)→限制性= 内切酶识别位点改变= (原来位点= ;消失或产生新的位点&#= 25110;= 识别位点= ;位移) →酶切电= 泳，片段<= span lang=3DZH-CN style=3D'font-family:SimSun;mso-fareast-font-family:SimSun; font-size:150%;mso-ansi-language:EN-US'>改变→确= 定是否存在致病基因= 3;

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限制性内切酶酶谱= 分析法举例

n镰形细胞贫ච= 0;原因：β-珠蛋白链第6位的GAG→GTG，造成Glu→Val所致。
染色体DNA

Bam HI

1.8kb= 片段
<= ![if !ppt]>
Hb19A

<= ![if !ppt]>
Hb19S

A

A：正&#= 24120;

B

B：ெ= 9;人

方法1

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限制性内切酶酶谱= 分析法举例 镰形细胞贫血=

染色体DNA

Mst I

电泳，印= ;迹，与β<= span lang=3DEN-US style=3D'font-size:89%;color:red;position:relative;top:-.45em; mso-text-raise:30%;mso-fareast-language:ZH-CN'>A或βS探针<= /span>分别杂交= ;

正常É= 54;

1.15<= /span>

kb=

1.15<= /span>

kb=

杂合é= 76;<= span lang=3DZH-CN style=3D'font-family:SimSun;mso-fareast-font-family:SimSun; font-size:117%;color:red;mso-ansi-language:EN-US'>病人<= /span>

纯合é= 76;<= span lang=3DZH-CN style=3D'font-family:SimSun;mso-fareast-font-family:SimSun; font-size:117%;color:red;mso-ansi-language:EN-US'>病人<= /span>

1.35<= /span>

kb=

1.35<= /span>

与βA杂ߝ= 2;

与βS杂&#= 20132;

与βS杂&#= 20132;

方法2

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限制性内切酶酶谱= 分析法举例 镰形细胞贫血=

294

191

103

bp

A

A：正&#= 24120;人

B

B：ಫ= 1;合子病人

C

C：੓= 4;合子病人

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=

2、DNA限制性片段长= 230;多态性分析

    在人类= 基因组中,平均约200对= 碱基可= 发生一对变异(称为中&#= 24615;突<= span lang=3DZH-CN style=3D'font-family:SimSun;mso-ascii-font-family:SimSun;mso-= fareast-font-family: SimSun;font-size:167%;mso-ansi-language:EN-US'>变),中ঝ= 5;突变导致个体间核苷&= #37240;序= 列的差= 异,称为DNA基因多&= #24577;性.

不少DNAä= 10;态性发生在限制性内= 切= 酶识别= 位点上,酶解该DNA片段就= ;会= 产生长= 度不同的片断,称为限࠸= 6;性= 片ૅ= 3;长度多态性.

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3、౔= 1;位基因特异寡核苷酸&= #25506;针杂交ૢ= 1;

=

=

n设计针对突ࡢ= 4;和正常的寡核苷酸探&= #38024;分别进行= ;杂交

n在严格条件ߍ= 9;进行杂交和洗膜

正常É= 54;

正常ÿ= 06;针

突变&= #25506;针

有突&= #21464;

正常ÿ= 06;针

突变&= #25506;针

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（二）聚合%= 238;链反应(PCR)

=

=

n扩增片段= 长度的变异：有插入= 或缺失时，扩增长度会增加或= 变小，与正常长度比= 6739;，找出致病基因

nPCR-RF= LP分析&#= 65306;相差1个碱基，= 改变酶切位点，<= span style=3D'position:absolute;top:35.29%;left:3.49%;width:96.5%;height:32.35%= '>因此经酶= 切后会产生正常基因= 5193;增的扩增产物长度不同的片= 段

nPCR-AS= O(SSO)分析&#= 65306;按严格条= 件进行杂交，找出致= 0149;基因。有多= 种探针可进行反向斑= 8857;杂交技术

nPCR-序列&#= 20998;析法：对于未知= 突变或少见的突变，= 1487;直接测序= 确定

n引物特异= 性PCR法：设计2对引物

nPCR-SS= CP分析&#= 27861;：有突变，= 长度不变，但构象改变，PAGE的迁移率= 改变

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=

（三）= 基因测序

    分离出患者௚= 0;有关基因，测定出 =

= 其碱基排列顺序，找= 0986;其变异所在。

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三= 289;基因诊断的应用

=

=

n诊断疾病：遗传病疾病ӌ= 9;多基因疾病、恶性肿<= /b>瘤和感染性ெ= 2;病

nHLA的基因分= 型：

n个体遗传基ࢰ= 0;的鉴定：DNA指纹分析、HLA基因分型、಩= 8;细胞血型基因检测、&= #21508;种DNA多态性分析

n结构基因及ࠥ= 4;基因表达的分析 <= /span>

n性别鉴定：性连锁遗传ெ= 9;分析、法医学的个人<= /b>鉴定、运动࡯= 2;的性别检查

n生物学分类ລ= 2;定

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第二节<= span lang=3DZH-CN style=3D'color:red;mso-ansi-language:EN-US'> 基因治疗

n一、基因= ;治疗的概念

n二、基因= ;治疗的基本程序

n三、基因= ;治疗的应用与展望 <= /span>

n

<= span style=3D'visibility:hidden'>n

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        将某种遗= ;传物质转移到患= = 者细胞内= ;，使其在体内发挥= 作用，以= ;达到治疗疾病目的= 的方法。 =

一、基因治疗的概= 念

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基因治疗= 340;基本方法

基因置换= ：将致病&#= 22522;因整个地以正常基Þ= 40;置换，使&#= 33268;病基因永久性得到ć= 56;正。

基因修正= ：纠正致病基ࢰ= 0;的突变碱基顺序，而将&#= 27491;常部分予以保留。 =

基因修饰= ：将目的基因ल= 8;入病变细胞或其他细胞，&#= 20197;目的基因的表达产Ĥ= 89;来补偿缺陷细胞的&#= 21151;能或使原来的功能ô= 71;到加强。

基因失活= ：应用反&#= 20041;技术特异封闭某些á= 22;因的表达&#= 65292;以抑制某些有害基Þ= 40;有表达

v

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二、基因= 835;疗的基本程序

n= （一）治= 0103;性基因的选择

n= （二）基= 2240;载体的选择

n= （三）靶= 2454;胞的选择

n= （四）基= 2240;转移

n= （五）外= 8304;基因表达的筛检

n= （六）回= 6755;体内

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         选择对= 疾病有治疗作用的= <= span lang=3DZH-CN style=3D'font-family:SimSun;mso-fareast-font-family:SimSun; font-size:200%;mso-ansi-language:EN-US'>特定目的= ;基因。

= （一）治疗性基因的= 6873;择

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获得&= #30446;的基因的方法

n基因的克= ;隆化

n基因的人= ;工合成

n基因的PCR扩增

n人体基因= ;组的降解

<= span style=3D'font-size:113%;visibility:hidden'>n

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    目前使用= ;的有病毒载体和非病&#= 27602;= 载体两大= ;类。基因治疗的临床&#= 23454;施<= span lang=3DZH-CN style=3D'font-family:SimSun;mso-fareast-font-family:SimSun; font-size:183%;mso-ansi-language:EN-US'>中，一般= ;多选用病毒载体。 <= /span>

        = ;

= （三）基因载体的选= 5321;

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载体的&= #36873;择

n= 物理法

n= 化学法

n= 融合法

表达载体

质粒载体= ;

(穿梭载体= ;)

病毒载体
细胞

导入

= 物理法

= 化学法

= 融合法

导入

细胞

包装

感染

靶细胞

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（三）靶= ;细胞的选择

     根据受体= ;细胞种类的不= 同，基因= ;治疗分为体细胞的= 基因治疗= ;和生殖细胞的基因= 治疗。

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受体细&= #32990;（靶细胞）的种类=

=

=

n淋巴细胞

n造血细胞

n上皮细胞

n角质细胞

n内皮细胞

n成纤维细胞= ;

n肝脏细胞

n肿瘤细胞

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（四）基= ;因转移

     将外源治= ;疗性基因导入受体细= 胞。

方法有两类：
1&= #12289;非病毒介导的基因$= 716;移；

2&= #12289;病毒介导的基因转= 227;

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将目的&= #22522;因导入靶细胞

n物理学方= ;法

n化学方法= ;

n融合法

<= span style=3D'font-size:113%;color:red;visibility:hidden'>n=

•病毒载体法（最好= ）

效率较低
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将目的&= #22522;因导入靶细胞

=

=

nDNA磷酸钙共沉ૹ= 6;法

n&= #26174;微注射法

n&= #22522;因枪(粒子轰击= 基因转移技术) =

n&= #30452;接注射法

n&= #30005;激转移

n&= #26579;色体介导转移法

n&= #21463;体介导转移法

n&= #36870;转录病毒介导转移= 861;

n&= #33146;病毒介导转移法

n&= #30168;苗病毒介导转移法 = ;

n&= #21516;源重组介导的靶向= 972;合(基因打靶= )

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= （五）外源基因表达= 0340;筛检        = ;

    在体外= 培养细胞中，基因转= 6579;率很= 难达到= 100%，需要利用载体中的= ;标记= 基因对= 转染细胞进行筛选。

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标&#= 35760;基因技术

n将目的基= ;因和标记基因重组在&#= 21516;一或不同表= ;达载体, 导入受体= ;细胞, 适时选用选= ;择性培养基, 筛选标记= ;基因表型, 使导入外= ;源基因的靶细胞存活,而未转导= ;的细胞则不能生存.

n如G418筛选

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G418筛选原理=

=

=

n新霉素是细= ;菌抗生素, 可干扰原核生物的= 蛋白质合成, 但对真核细胞则无= 影响。

nG418是新霉素的౲= 7;似物，对原核和真核&= #32454;胞均有毒性Ӎ= 0;

n细菌的新霉素磷%= 240;转移酶(neo)基因在真核细胞中表达, 当neo基因与能有= ;效转录的真核DNA序列连锁时就能获= 得有效有表达。

n产生的新霉= ;素磷酸转移酶可使G418失活。

n当真核细胞= ;转入neo基因后，就ߩ= 0;含G418的选择性培养基中存= 活，而未转入neo基因的细胞则不能ஶ= 3;存。

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（六）回= ;输体内

    将治疗性基ࢰ= 0;修饰的细胞以不同= 的方式回= ;输体内以发挥治疗效&#= 26524;。

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三、基因= 835;疗的应用与展望

n基因治疗= ;的构想阶段（1953<= /span>～）

n基因治疗= ;的实验阶段（1980 = ～）

n基因治疗= ;的临床应用阶段= （1989.1.19= ～; 国内1991.12.3= 1开始血友病的= ;基因治疗)

n基因治疗= ;的意义和展望
<= span style=3D'font-size:113%;visibility:hidden'>n

<= span style=3D'visibility:hidden'>n

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人类= ;基因治疗的意义

=

=

n免疫调节: 将抗= 407;的基因或抗体经基因= ;治疗途径导= 837;人体<= b>,以改= 464;人体免疫反应应答和= ;调节<= b>,对疾= 149;的防治有重要意义<= b>. 将细 = 990;因子基因导入人= 307;进行抗病毒和抗肿瘤= ;的基因治疗是目前基因= 835;疗研究非常活跃的领= ;域。

n遗传缺陷的纠正：= 第一次为基因缺陷的= 6951;传病提供= 102;治愈的机会

n分泌治疗性的蛋白= 质，发挥治疗作用

n封闭有害基因的表= 达：以反义技术或反= 0041;失活效应= 233;制、封闭肿瘤基因和= ;病毒基因，治疗病毒性= 644;肿瘤性疾病。

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实现基&= #22240;治疗面临的困难<= /div>

n基因缺陷= ;病的早期诊断还很困&#= 38590;

n基因载体= ;本身以及外源性基因&#= 25972;合到宿主细胞染= ;色体的位点多是随机&#= 30340;

n导入的基= ;因在宿主细胞内的表&#= 36798;水平难于控制，= ;表达也有一定的随机&#= 24615;

n对于种类= ;更多的、病人数量更&#= 22823;和遗传易感性疾= ;病其基因治疗的难度&#= 26356;大

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基因= 7835;疗的前景

n前景&= #20809;明

n任重&= #32780;道远

n21世纪&= #30340;人类基因治疗，就= 687;20世纪的&= #20813;疫预防和抗生素的= 212;用，将给人&= #31867;的健康事业带来极= 854;深刻的影响<= /b>

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页眉

‹日期/时间›

单击此处编= 6753;母版文本样式

第二级

第三级

第四级

第五级

‹= 页脚›

‹#›

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YW50O30NCmE6YWN0aXZlDQoJe2NvbG9yOiNGQUMxNjQgIWltcG9ydGFudDt9DQphOnZpc2l0ZWQN Cgl7Y29sb3I6I0MzRTY4NCAhaW1wb3J0YW50O30NCn== ------=_NextPart_01C48438.3E875CB0 Content-Location: file:///C:/85899A72/22.files/script.js Content-Transfer-Encoding: quoted-printable Content-Type: application/javascript; charset="us-ascii" function LoadSld() { var sld=3DGetObj("SlideObj") if( !g_supportsPPTHTML ) { =09 sld.style.visibility=3D"visible" return } if( MakeNotesVis() ) return runAnimations =3D _InitAnimations(); =09 if( IsWin("PPTSld") ) parent.SldUpdated(GetSldId()) g_origSz=3DparseInt(SlideObj.style.fontSize) g_origH=3Dsld.style.posHeight g_origW=3Dsld.style.posWidth g_scaleHyperlinks=3D(document.all.tags("AREA").length>0) if( g_scaleHyperlinks ) InitHLinkArray() if( g_scaleInFrame||(IsWin("PPTSld") && parent.IsFullScrMode() ) ) document.body.scroll=3D"no" _RSW() if( IsWin("PPTSld") && parent.IsFullScrMode() ) FullScrInit(); =09 MakeSldVis(); ChkAutoAdv() if( runAnimations ) { if( document.all("NSPlay") ) document.all("NSPlay").autoStart =3D false; if( sld.filters && sld.filters.revealtrans ) setTimeout( "document.body.start()", sld.filters.revealtrans.duration * = 1000 ); else document.body.start(); } } function MakeSldVis()=20 { var fTrans=3Dg_showAnimation && SldHasTrans() if( fTrans )=09 { if( g_bgSound ) { idx=3Dg_bgSound.indexOf(","); pptSound.src=3Dg_bgSound.substr( 0, idx ); pptSound.loop=3D -(parseInt(g_bgSound.substr(idx+1))); } SlideObj.filters.revealtrans.Apply()=09 } SlideObj.style.visibility=3D"visible" if( fTrans ) SlideObj.filters.revealtrans.Play() } function MakeNotesVis()=20 { if( !IsNts() ) return false=20 SlideObj.style.display=3D"none" nObj =3D document.all.item("NotesObj") parent.SetHasNts(0) if( nObj ) {=20 nObj.style.display=3D"" parent.SetHasNts(1) } return 1 } function ChkAutoAdv() { if(SldHasTrans()) SlideObj.onfilterchange=3DAutoAdv else AutoAdv() } function AutoAdv() { if(!IsWin("PPTSld") || !gUseSldTimings )return var sld=3DGetCurSld() if( (sld.mAdvDelay>0) && !parent.IsFramesMode() ) setTimeout("parent.GoToNextSld()",sld.mAdvDelay) } function GetObj(id) { if(g_supportsPPTHTML) return document.all(id); else return document.getElementById(id); } function SldHasTrans() { return SlideObj.style.filter !=3D ""; } function GetSldId() { return sId=3Dlocation.href.substring(location.href.la= stIndexOf('/')+1) } function HideMenu() { if( frames["PPTSld"] && PPTSld.document.all.item("ctx= tmenu") && PPTSld.ctxtmenu.style.display!=3D"none" ) { PPTSld.ctxtmenu.styl= e.display=3D'none'; return true } return false } function IsWin( name ) { return window.name =3D=3D name } function IsNts() { return IsWin("PPTNts") } function IsSldOrNts() { return( IsWin("PPTSld")||IsWin("PPTNts") ) } function SupportsPPTAnimation() { return( navigator.platform =3D=3D "Win32"= && navigator.appVersion.indexOf("Windows")>0 ) } function SupportsPPTHTML() { var appVer=3Dnavigator.appVersion, msie=3DappVer.indexOf("MSIE "), ver=3D0 if( msie >=3D 0 ) ver=3DparseFloat( appVer.substring( msie+5, appVer.indexOf(";",msie) ) ) else ver=3DparseInt(appVer) return( ver >=3D 4 && msie >=3D 0 ) } function _RSW() { if( !g_supportsPPTHTML || IsNts() || ( !g_scaleInFrame && (!IsWin("PPTSld") || !parent.IsFullScrMode()) ) ) return var padding=3D0; if( IsWin("PPTSld") && parent.IsFramesMode() ) padding=3D6 cltWidth=3Ddocument.body.clientWidth-padding cltHeight=3Ddocument.body.clientHeight-padding factor=3D(1.0*cltWidth)/g_origW if( cltHeight < g_origH*factor ) factor=3D(1.0*cltHeight)/g_origH newSize =3D g_origSz * factor if( newSize < 1 ) newSize=3D1 s=3DSlideObj.style s.fontSize=3DnewSize+"px" s.posWidth=3Dg_origW*factor s.posHeight=3Dg_origH*factor s.posLeft=3D(cltWidth-s.posWidth+padding)/2 s.posTop=3D(cltHeight-s.posHeight+padding)/2 if( g_scaleHyperlinks ) ScaleHyperlinks( factor ) } function _InitAnimations() { animRuntimeInstalled =3D ''+document.body.localTime !=3D 'undefined'; isFullScreen =3D (window.name =3D=3D "PPTSld") && !parent.IsFramesMode(); g_animUseRuntime =3D g_showAnimation && animRuntimeInstalled && !(isFullSc= reen && parent.IsSldVisited()); if( g_animUseRuntime ) { collSeq =3D document.all.tags("seq"); if( collSeq !=3D null ) { for(ii=3D0;ii numSlds ) gSldJumpIdx =3D numSlds; if ( gSldJumpIdx >=3D 0 ) { if ( gSldJumpIdx =3D=3D 0 ) gSldJumpIdx =3D 1; var jumpTo =3D parseInt(gSldJumpIdx); gSldJump =3D 0; gSldJumpIdx =3D ""; win.GoToSld( parent.GetSldList().mList[jumpTo-1].mSldHref ) } } } function _KDH() { if( event.keyCode =3D=3D 8 ) { event.returnValue =3D 0; parent.GoToPrevSld(); } }function DocumentOnClick() { if( IsNts() || parent.HideMenu() ) return; if( ( g_allowAdvOnClick && (window.name=3D=3D"PPTSld") && !parent.IsFrames= Mode() ) || (event && event.keyCode=3D=3D32) ) { =09 if( g_animUseRuntime && g_animMainSequence && g_animMainSequence.cangonex= t ) return; parent.GoToNextSld(); } } var g_supportsPPTHTML =3D SupportsPPTHTML(), g_scaleInFrame =3D 1, gId=3D""= , g_bgSound=3D"", g_scaleHyperlinks =3D false, g_allowAdvOnClick =3D 1, g_showInBrowser = =3D 0, gLoopCont =3D 0, gUseSldTimings =3D 1; var g_showAnimation =3D g_supportsPPTHTML && SupportsPPTAnimation() && ( (w= indow.name=3D=3D"PPTSld" && !parent.IsFramesMode()) || g_showInBrowser );va= r g_animManager =3D null; var g_animUseRuntime =3D false; var g_animItemsToHide, g_animInteractiveItems, g_animSlideTime; var g_animMainSequence =3D null; var ENDSHOW_MESG=3D"放映结束，单击= ;鼠标退出。", SCREEN_MODE=3D"Frames", gIsEndS= how=3D0, NUM_VIS_SLDS=3D36, SCRIPT_HREF=3D"script.js", FULLSCR_HREF=3D"full= screen.htm"; var gCurSld =3D gPrevSld =3D 1, g_offset =3D 0, gNtsOpen =3D gHasNts =3D gO= tlTxtExp =3D 0, gHasNarration =3D 0, gOtlOpen =3D true window.gPPTHTML=3DSupportsPPTHTML() var gMainDoc=3Dnew Array(new hrefList("slide0001.htm",1,-1,1),new hrefList(= "slide0002.htm",1,-1,1),new hrefList("slide0003.htm",1,-1,1),new hrefList("= slide0004.htm",1,-1,1),new hrefList("slide0005.htm",1,-1,1),new hrefList("s= lide0006.htm",1,-1,1),new hrefList("slide0026.htm",1,-1,1),new hrefList("sl= ide0045.htm",1,-1,1),new hrefList("slide0046.htm",1,-1,1),new hrefList("sli= de0047.htm",1,-1,1),new hrefList("slide0030.htm",1,-1,1),new hrefList("slid= e0048.htm",1,-1,1),new hrefList("slide0049.htm",1,-1,1),new hrefList("slide= 0033.htm",1,-1,1),new hrefList("slide0050.htm",1,-1,1),new hrefList("slide0= 008.htm",1,-1,1),new hrefList("slide0016.htm",1,-1,1),new hrefList("slide00= 17.htm",1,-1,1),new hrefList("slide0009.htm",1,-1,1),new hrefList("slide001= 8.htm",1,-1,1),new hrefList("slide0034.htm",1,-1,1),new hrefList("slide0019= .htm",1,-1,1),new hrefList("slide0035.htm",1,-1,1),new hrefList("slide0020.= htm",1,-1,1),new hrefList("slide0036.htm",1,-1,1),new hrefList("slide0024.h= tm",1,-1,1),new hrefList("slide0037.htm",1,-1,1),new hrefList("slide0038.ht= m",1,-1,1),new hrefList("slide0021.htm",1,-1,1),new hrefList("slide0039.htm= ",1,-1,1),new hrefList("slide0040.htm",1,-1,1),new hrefList("slide0022.htm"= ,1,-1,1),new hrefList("slide0010.htm",1,-1,1),new hrefList("slide0042.htm",= 1,-1,1),new hrefList("slide0043.htm",1,-1,1),new hrefList("slide0044.htm",1= ,-1,1)); /********************************************* Frameset functions These functions control slide navigation and state of the frameset. **********************************************/ function FullScrInit() { g_allowAdvOnClick =3D GetCurSld().mAdvOnClk document.body.style.backgroundColor=3D"black" document.oncontextmenu=3Dparent._CM; document.onkeydown =3D _KDH; document.ondragstart=3DCancel document.onselectstart=3DCancel self.focus() } function Redirect( frmId ) {=09 var str=3Ddocument.location.hash,idx=3Dstr.indexOf('#'), sId=3DGetSldId() if(idx>=3D0) str=3Dstr.substr(1); if( window.name !=3D frmId && ( sId !=3D str) ) { obj =3D GetObj("Main-File") window.location.href=3Dobj.href+"#"+sId return 1 } return 0 } var MHTMLPrefix =3D CalculateMHTMLPrefix();=20 function CalculateMHTMLPrefix() { if ( document.location.protocol =3D=3D 'mhtml:') {=20 href=3Dnew String(document.location.href)=20 Start=3Dhref.indexOf('!')+1=20 End=3Dhref.lastIndexOf('/')+1=20 if (End < Start)=20 return href.substring(0, Start)=20 else=20 return href.substring(0, End)=20 } return ''; } function GetTags(base,tag) { if(g_supportsPPTHTML) return base.all.tags(tag); else return base.getElementsByTagName(tag); } function UpdNtsPane(){ if(frames["PPTNts"]) PPTNts.location.replace( MHTMLP= refix+GetHrefObj( gCurSld ).mNtsHref ) } function UpdNavPane( sldIndex ){ if(gNavLoaded) PPTNav.UpdNav() } function UpdOtNavPane(){ if(gOtlNavLoaded) PPTOtlNav.UpdOtlNav() } function UpdOtlPane(){ if(gOtlLoaded) PPTOtl.UpdOtl() } function SetHasNts( fVal ) { if( gHasNts !=3D fVal ) { gHasNts=3DfVal UpdNavPane() } } function ToggleOtlText() { gOtlTxtExp=3D!gOtlTxtExp UpdOtlPane() } function ClearMedia() { // Clear any sounds playing before launching another browser window. Other= wise, // in fullscreen mode, you'll continue to hear the sound in the frames mod= e. if (PPTSld.pptSound) PPTSld.pptSound.loop =3D 0; } function FullScreen() {=20 if ( PPTSld.g_animUseRuntime ) PPTSld.document.body.pause(); ClearMedia(); var href =3D ( document.location.protocol =3D=3D 'mhtml:') ? FULLSCR_HREF = : FULLSCR_HREF+"#"+GetHrefObj(gCurSld).mSldHref; if(PPTNav.event.ctrlKey) { var w =3D (window.screen.availWidth * 1.0) / 2.0 var h =3D w * (PPTSld.g_origH * 1.0) / PPTSld.g_origW win =3D window.open( MHTMLPrefix+href,null,"toolbar=3D0,resizable=3D1,top= =3D0,left=3D0," + "width=3D"+ w + ",height=3D" + h ); if( win.document.body && PPTSld.g_animUseRuntime ) win.document.body.PPTSldFrameset=3Dwindow; } else { win =3D window.open( MHTMLPrefix+href,null,"fullscreen=3Dyes" ); if( win.document.body && PPTSld.g_animUseRuntime ) win.document.body.PPTSldFrameset=3Dwindow; } } function ToggleVNarration() { rObj=3DPPTSld.document.all("NSPlay") if( rObj && !PPTSld.g_animUseRuntime ) { if( (rObj.playState =3D=3D 1)||(rObj.playState =3D=3D 0) ) rObj.Play() else if( rObj.playState =3D=3D 2 ) rObj.Pause() else return; } else if( PPTSld.g_animUseRuntime ) { narObj =3D PPTSld.document.all("narrationID") if( narObj ) narObj.togglePause() } } function GetCurSldNum() { =20 obj=3DGetHrefObj(gCurSld) if( obj.mOrigVis =3D=3D 1 ) return obj.mSldIdx else =20 return gCurSld } function GetNumSlds() { =20 if( GetHrefObj(gCurSld).mOrigVis =3D=3D 1 ) return GetSldList().mNumVisSlds; else return GetSldList().mList.length } function GetSldNum( href ) { for(ii=3D0; ii 1 ) PopSldList(); else if( !IsFramesMode() ) { if( gLoopCont ) GoToFirst() else EndShow() } } function GoToPrevSld() { ii=3DgCurSld-1 if( ii > 0 ) { obj=3DGetHrefObj(ii) while ( obj && ( obj.mVis =3D=3D 0 ) && ( ii>0 ) ) obj=3DGetHrefObj(--ii) if( ii =3D=3D 0 ) ii=3D1 GoToSldNum(ii) } } function GoToFirst(){ GoToSld( GetHrefObj(1).mSldHref ) } function GoToLast() { ii=3DGetSldList().mList.length if( ii !=3D gCurSld ) GoToSld( GetHrefObj(ii).mSldHref ) } function GoToSldNum( num ) { if( PPTSld.event ) PPTSld.event.cancelBubble=3Dtrue obj =3D GetHrefObj( num ) obj.mVis=3D1 gPrevSld=3DgCurSld gCurSld =3D num; PPTSld.location.replace(MHTMLPrefix+obj.mSldHref) if( IsFramesMode() ) { UpdNavPane(); UpdOtlPane(); UpdNtsPane() } } function GoToSld( href ) { if( PPTSld.event ) PPTSld.event.cancelBubble=3Dtrue GetHrefObj( GetSldNum(href) ).mVis=3D1 PPTSld.location.replace(MHTMLPrefix+href) } function SldUpdated( id ) { if( id =3D=3D GetHrefObj(gCurSld).mSldHref ) return gPrevSld=3DgCurSld gCurSld=3DGetSldNum(id) if( IsFramesMode() ) { UpdNavPane(); UpdOtlPane(); UpdNtsPane() } } function PrevSldViewed(){ GoToSld( GetHrefObj(gPrevSld).mSldHref ) } function HasPrevSld() { return ( gIsEndShow || ( gCurSld !=3D 1 && GetHrefO= bj( gCurSld-1 ).mVis =3D=3D 1 )||( GetCurSldNum() > 1 ) ) } function HasNextSld() { return (GetCurSldNum() !=3D GetNumSlds()) } function CloseWindow() { if( HideMenu() ) return; =09 var event =3D PPTSld.event; if( !IsFramesMode() && event && (event.keyCode=3D=3D27 || event.keyCode=3D= =3D32 || event.type=3D=3D"click" ) ) window.close( self ); CatchNumKeys( self, event ); } function Unload() { gIsEndShow=3D0; } function SetupEndShow() { gIsEndShow=3D1; PPTSld.document.body.scroll=3D"no"; PPTSld.document.onkeypress=3DCloseWindow; PPTSld.document.onclick=3DCloseWindow; PPTSld.document.oncontextmenu=3D_CM; } function EndShow() { if( IsFramesMode() ) return if( PPTSld.event ) PPTSld.event.cancelBubble=3Dtrue doc=3DPPTSld.document var dir =3D doc.body.dir if( dir !=3D "rtl" ) dir =3D "ltr"; doc.open() doc.writeln('

' + ENDSHOW_MESG + '
') doc.close() } function SetSldVisited(){ GetSldList().mList[gCurSld-1].mVisited=3Dtrue } function IsSldVisited(){ return GetSldList().mList[gCurSld-1].mVisited } function hrefList( sldHref, visible, advDelay, advClk ) { this.mSldHref=3D this.mNtsHref =3D sldHref this.mOrigVis=3D this.mVis =3D visible this.mVisited=3D false this.mAdvDelay=3D advDelay this.mAdvOnClk=3D advClk } function SldList(arr,curSld,fEnd) { this.mCurSld =3D curSld; this.mList =3D new Array(); var idx =3D 1; for(ii=3D0;ii 0) { PushSldList(sldList,fEnd); gCurSld =3D 1; } else if( PPTSld.event ) PPTSld.event.cancelBubble=3Dtrue } function PushSldList(arr,fEnd) { var ii =3D gSldStack.length; gSldStack[ii] =3D new SldList(arr,gCurSld,fEnd); GoToSld( gSldStack[ii].mList[0].mSldHref ); } function PopSldList() { if (gSldStack[gSldStack.length-1].fEndShow) EndShow() else { gCurSld =3D gSldStack[gSldStack.length-1].mCurSld; gSldStack[gSldStack.length-1] =3D null; gSldStack.length--; var sldList =3D gSldStack[gSldStack.length-1]; GoToSld( sldList.mList[gCurSld - 1].mSldHref ); } } var custShowList=3Dnew Array(); /********************************************* Navigation button implementation There are 2 types of buttons: ImgBtn, TxtBtn implemented as function objects. They share a similiar interface so the event handlers can call SetActive, for example, on a button=20 object without needing to know exactly=20 what type of button it is. **********************************************/ //---------------------------------- function ImgBtn( oId,bId,w,action ) //---------------------------------- { var t=3Dthis t.Perform =3D _IBP t.SetActive =3D _IBSetA t.SetInactive=3D _IBSetI t.SetPressed =3D _IBSetP t.SetDisabled=3D _IBSetD t.Enabled =3D _IBSetE t.ChangeIcon =3D null t.UserAction =3D action t.ChgState =3D _IBUI t.mObjId =3D oId t.mBorderId=3D bId t.mWidth =3D w t.mIsOn =3D t.mCurState =3D 0 } function _IBSetA() { if( this.mIsOn ) { obj=3Dthis.ChgState( gHiliteClr,gShadowClr,2 ) obj.style.posTop=3D0 } } function _IBSetI() { if( this.mIsOn ) { obj=3Dthis.ChgState( gFaceClr,gFaceClr,1 ) obj.style.posTop=3D0=20 } } function _IBSetP() { if( this.mIsOn ) { obj=3Dthis.ChgState( gShadowClr,gHiliteClr,2 ) obj.style.posLeft+=3D1; obj.style.posTop+=3D1 } } function _IBSetD() { =20 obj=3Dthis.ChgState( gFaceClr,gFaceClr,0 ) obj.style.posTop=3D0=20 } function _IBSetE( state ) { var t=3Dthis GetObj( t.mBorderId ).style.visibility=3D"visible" if( state !=3D t.mIsOn ) { t.mIsOn=3Dstate if( state ) t.SetInactive() else t.SetDisabled() } } function _IBP() { var t=3Dthis if( t.mIsOn ) { if( t.UserAction !=3D null ) t.UserAction() if( t.ChangeIcon ) { obj=3DGetObj(t.mObjId) if( t.ChangeIcon() ) obj.style.posLeft=3Dobj.style.posLeft+(t.mCurState-4)*t.mWidth else obj.style.posLeft=3Dobj.style.posLeft+(t.mCurState-0)*t.mWidth } t.SetActive() } =20 } function _IBUI( clr1,clr2,nextState ) { var t=3Dthis SetBorder( GetObj( t.mBorderId ),clr1,clr2 ) obj=3DGetObj( t.mObjId ) obj.style.posLeft=3Dobj.style.posLeft+(t.mCurState-nextState)*t.mWidth-obj= .style.posTop t.mCurState=3DnextState return obj } //----------------------------------------- function TxtBtn( oId,oeId,action,chkState ) //----------------------------------------- { var t=3Dthis t.Perform =3D _TBP t.SetActive =3D _TBSetA t.SetInactive=3D _TBSetI t.SetPressed =3D _TBSetP t.SetDisabled=3D _TBSetD t.SetEnabled =3D _TBSetE t.GetState =3D chkState t.UserAction =3D action t.ChgState =3D _TBUI t.mObjId =3D oId t.m_elementsId=3D oeId t.mIsOn =3D 1 } function _TBSetA() { var t=3Dthis if( t.mIsOn && !t.GetState() ) t.ChgState( gHiliteClr,gShadowClr,0,0 ) } function _TBSetI() { var t=3Dthis if( t.mIsOn && !t.GetState() ) t.ChgState( gFaceClr,gFaceClr,0,0 ) } function _TBSetP() { if( this.mIsOn ) this.ChgState( gShadowClr,gHiliteClr,1,1 ) } function _TBSetD() { =20 this.ChgState( gFaceClr,gFaceClr,0,0 ) this.mIsOn =3D 0 } function _TBSetE() { var t=3Dthis if( !t.GetState() ) t.ChgState( gFaceClr,gFaceClr,0,0 ) else t.ChgState( gShadowClr,gHiliteClr,1,1 ) t.mIsOn =3D 1 } function _TBP() { var t=3Dthis if( t.mIsOn ) {=20 if( t.UserAction !=3D null ) t.UserAction() if( !t.GetState ) return if( t.GetState() ) t.SetPressed() else t.SetActive() } =20 } function _TBUI( clr1,clr2,lOffset,tOffset ) { SetBorder( GetObj( this.mObjId ),clr1,clr2 ) Offset( GetObj( this.m_elementsId ),lOffset,tOffset ) } function Offset( obj, top, left ){ obj.style.top=3Dtop; obj.style.left=3Dle= ft } function SetBorder( obj, upperLeft, lowerRight ) { s=3Dobj.style; s.borderStyle =3D "solid" s.borderWidth =3D 1=20 s.borderLeftColor =3D s.borderTopColor =3D upperLeft s.borderBottomColor=3D s.borderRightColor =3D lowerRight } function GetBtnObj(){ return gBtnArr[window.event.srcElement.id] } function BtnOnOver(){ b=3DGetBtnObj(); if( b !=3D null ) b.SetActive() } function BtnOnDown(){ b=3DGetBtnObj(); if( b !=3D null ) b.SetPressed() } function BtnOnOut(){ b=3DGetBtnObj(); if( b !=3D null ) b.SetInactive() } function BtnOnUp() { b=3DGetBtnObj() if( b !=3D null ) b.Perform() else Upd() } function GetNtsState(){ return parent.gNtsOpen } function GetOtlState(){ return parent.gOtlOpen } function GetOtlTxtState(){ return parent.gOtlTxtExp } function NtsBtnSetFlag( fVal ) { s=3Ddocument.all.item( this.m_flagId ).style s.display=3D"none" if( fVal ) s.display=3D"" else s.display=3D"none" } function _BSetA_Border(){ b =3D gBtnArr[this.mObjId]; if( b !=3D null ) b.S= etActive() } function _BSetI_Border(){ b =3D gBtnArr[this.mObjId]; if( b !=3D null ) b.S= etInactive() } function _BSetP_Border(){ b =3D gBtnArr[this.mObjId]; if( b !=3D null ) b.S= etPressed() } function _BSetA_BorderImg() {=20 b =3D gBtnArr[this.mBorderId]=20 if( b !=3D null && this.mIsOn && !b.GetState() ) { obj=3Dthis.ChgState( gHiliteClr,gShadowClr,2 ) obj.style.posTop=3D0 } } function _BSetI_BorderImg() {=20 b =3D gBtnArr[this.mBorderId] if( b !=3D null && this.mIsOn && !b.GetState() ) { obj=3Dthis.ChgState( gFaceClr,gFaceClr,1 ) obj.style.posTop=3D0 } } var gHiliteClr=3D"THREEDHIGHLIGHT",gShadowClr=3D"THREEDSHADOW",gFaceClr=3D"= THREEDFACE" var gBtnArr =3D new Array() gBtnArr["nb_otl"] =3D new TxtBtn( "nb_otl","nb_otlElem",parent.ToggleOtlPan= e,GetOtlState ) gBtnArr["nb_otlElem"] =3D new TxtBtn( "nb_otl","nb_otlElem",parent.ToggleOt= lPane,GetOtlState ) gBtnArr["nb_nts"] =3D new ImgBtn( "nb_nts","nb_ntsBorder",10,parent.ToggleN= tsPane ) gBtnArr["nb_nts"].SetActive =3D _BSetA_BorderImg; gBtnArr["nb_nts"].SetInactive =3D _BSetI_BorderImg; gBtnArr["nb_ntsBorder"] =3D new TxtBtn( "nb_ntsBorder","nb_ntsElem",parent.= ToggleNtsPane,GetNtsState ) gBtnArr["nb_ntsElem"] =3D new TxtBtn( "nb_ntsBorder","nb_ntsElem",parent.To= ggleNtsPane,GetNtsState ) gBtnArr["nb_prevBorder"] =3D gBtnArr["nb_prev"]=3D new ImgBtn( "nb_prev","n= b_prevBorder",30,parent.GoToPrevSld ) gBtnArr["nb_nextBorder"] =3D gBtnArr["nb_next"]=3D new ImgBtn( "nb_next","n= b_nextBorder",30,parent.GoToNextSld ) gBtnArr["nb_sldshw"]=3D new ImgBtn( "nb_sldshw","nb_sldshwBorder",18,parent= .FullScreen ) gBtnArr["nb_sldshwBorder"] =3D new TxtBtn( "nb_sldshw","nb_sldshwBorder",pa= rent.FullScreen,null ) gBtnArr["nb_sldshwBorder"].SetActive =3D _BSetA_Border; gBtnArr["nb_sldshwBorder"].SetInactive =3D _BSetI_Border; gBtnArr["nb_sldshwText"] =3D new TxtBtn( "nb_sldshw","nb_sldshwText",parent= .FullScreen,null ) gBtnArr["nb_sldshwText"].SetActive =3D _BSetA_Border; gBtnArr["nb_sldshwText"].SetInactive =3D _BSetI_Border; gBtnArr["nb_voice"] =3D gBtnArr["nb_voiceBorder"] =3D new ImgBtn( "nb_voice= ","nb_voiceBorder",18,parent.ToggleVNarration ) gBtnArr["nb_otlTxtBorder"] =3D gBtnArr["nb_otlTxt"]=3D new ImgBtn( "nb_otlT= xt","nb_otlTxtBorder",23,parent.ToggleOtlText ) gBtnArr["nb_ntsBorder"].m_flagId=3D "nb_nts" gBtnArr["nb_ntsBorder"].SetFlag =3D NtsBtnSetFlag gBtnArr["nb_otlTxt"].ChangeIcon=3D GetOtlTxtState /********************************************* Context menu implementation _CM() is the function that's hooked up to the oncontextmenu event. Once we're asked to show the menu, we first build it by creating DIVs on-the-fly. Then we position it=20 within the screen area so it doesn't get clipped. Creating the DIVs using createElement() means we don't have to write out any extra HTML into the slide HTML files. **********************************************/ var sNext=3D"下一步",sPrev=3D"上一步",s= End=3D"结束放映",sFont=3D"宋体",sArrow= =3D"箭头",sFreeform=3D"任意形状",sRect= =3D"矩形",sOval=3D"椭圆" function ShowMenu() { BuildMenu(); var doc=3DPPTSld.document.body,x=3DPPTSld.event.clientX+doc.scrollLeft,y= =3DPPTSld.event.clientY+doc.scrollTop m =3D PPTSld.document.all.item("ctxtmenu") m.style.pixelLeft=3Dx if( (x+m.scrollWidth > doc.clientWidth)&&(x-m.scrollWidth > 0) ) m.style.pixelLeft=3Dx-m.scrollWidth m.style.pixelTop=3Dy if( (y+m.scrollHeight > doc.clientHeight)&&(y-m.scrollHeight > 0) ) m.style.pixelTop=3Dy-m.scrollHeight m.style.display=3D"" } function _CM() { if( !parent.IsFullScrMode() ) return; if(!PPTSld.event.ctrlKey) { ShowMenu() return false } else HideMenu() } function BuildMenu() { if( PPTSld.document.all.item("ctxtmenu") ) return var mObj=3DCreateItem( PPTSld.document.body ) mObj.id=3D"ctxtmenu" mObj.style.visibility=3D"hidden" var s=3DmObj.style s.position=3D"absolute" s.cursor=3D"default" s.width=3D"120px" SetCMBorder(mObj,"menu","black") var iObj=3DCreateItem( mObj ) SetCMBorder( iObj, "threedhighlight","threedshadow" ) iObj.style.padding=3D2 CreateMenuItem( iObj,sNext,M_GoNextSld,M_True ) CreateMenuItem( iObj,sPrev,M_GoPrevSld,M_HasPrevSld ) =09 CreateSeparator( iObj ) CreateMenuItem( iObj,sEnd,M_End,M_True ) mObj.style.visibility=3D"visible" } function Cancel() { window.event.cancelBubble=3Dtrue; window.event.returnVa= lue=3Dfalse } function Highlight() { ChangeClr("activecaption","threedhighlight") } function Deselect() { ChangeClr("threedface","menutext") } function Perform() { e=3DPPTSld.event.srcElement if( e.type=3D=3D"menuitem" && e.IsActive() ) e.Action() else PPTSld.event.cancelBubble=3Dtrue } function ChangeClr( bg,clr ) { e=3DPPTSld.event.srcElement if( e.type=3D=3D"menuitem" && e.IsActive() ) { e.style.backgroundColor=3Dbg e.style.color=3Dclr } } function M_HasPrevSld() { return( parent.HasPrevSld() ) } function M_GoNextSld() { if( gIsEndShow ) M_End(); else GoToNextSld() } function M_GoPrevSld() { if( gIsEndShow ) { gIsEndShow=3D0; history.back();= PPTSld.event.cancelBubble=3Dtrue; } else GoToPrevSld() } function M_True() { return true } function M_End() { window.close( self ) } function CreateMenuItem( node,text,action,eval ) { var e=3DCreateItem( node ) e.type=3D"menuitem" e.Action=3Daction e.IsActive=3Deval e.innerHTML=3Dtext if( !e.IsActive() ) e.style.color=3D"threedshadow" e.onclick=3DPerform e.onmouseover=3DHighlight e.onmouseout=3DDeselect s=3De.style; s.fontFamily=3DsFont s.fontSize=3D"9pt" s.paddingLeft=3D2 } function CreateSeparator( node ) { var sObj=3DCreateItem( node ) SetCMBorder(sObj,"menu","menu") var s=3DsObj.style s.borderTopColor=3D"threedshadow" s.borderBottomColor=3D"threedhighlight" s.height=3D1 s.fontSize=3D"0px" } function CreateItem( node ) { var elem=3DPPTSld.document.createElement("DIV") node.insertBefore( elem ) return elem } function SetCMBorder( o,ltClr,rbClr ) { var s=3Do.style s.backgroundColor=3D"menu" s.borderStyle=3D"solid" s.borderWidth=3D1 s.borderColor=3DltClr+" "+rbClr+" "+rbClr+" "+ltClr } ------=_NextPart_01C48438.3E875CB0 Content-Location: file:///C:/85899A72/22.files/fullscreen.htm Content-Transfer-Encoding: quoted-printable Content-Type: text/html; charset="us-ascii"
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&#= 22791;注

大纲

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第一= 节基因诊断=

一、基因诊断的概= 565;和特点

二、基因诊断的常= 992;技术方法

三、基因诊断的应= 992;

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（一= ）基因诊断的概念: 利用现代分子生物学&= #25216;术和分子遗传学的= 216;术方法，直接检测基= ;因结构及其表达水平&#= 26159;否正常，从而对疾ĭ= 49;作出诊断的方法。

一、基因诊断的概= 565;和特点

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（二= ）基因诊断的特点=

针对性强

特异性高

灵敏度高

适用性强

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二、= 基因诊断的常用技术= 6041;法

（一）、核酸分子= 434;交技术

（二）、聚合酶链= 453;应(PCR)

（三）、基因测序

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（一= ）核酸分子杂交技术

限制性内切酶酶谱= 998;析法

DNA限制性片段长度多&= #24577;性分析

等位基因特异寡核!= 527;酸探针杂交法

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限制= 性内切酶酶谱分析法= 0030;例

镰形细胞贫血原因A= 306;β-珠蛋白链第6位的GAGU= 94;GTG，造成Glu→Val所致。

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限制= 性内切酶酶谱分析法= 0030;例
镰形细胞贫血

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限制= ;性内切酶酶谱分析法&#= 20030;例
镰形细胞贫血

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3、౔= 1;位基因特异寡核苷酸&= #25506;针杂交法

设计针对突变和正= 120;的寡核苷酸探针分别= ;进行杂交

在严格条件下进行= 434;交和洗膜

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（二= ;）聚合酶链反应(PCR)

扩增片段长度的变= 322;：有插入或缺失时，= ;扩增长度会增加或变&#= 23567;，与正常长度比较ʌ= 92;找出致病基因

PCR-RFLP分析：相差1个碱= 2522;，改变酶切位点，ࢰ= 0;此经酶切后会产生正&= #24120;基因扩增的扩增产= 289;长度不同的片段

PCR-ASO(SSO)分析：按严格条= 件进行杂交，找出致= 0149;基因。有多种探针ࡤ= 7;进行反向斑点杂交技&= #26415;

PCR-序列分析法：对于= 未知突变或少见的突= 1464;，可直接测序确定

引物特异性PCR法：设&= #35745;2对引物

PCR-SSCP分析法：有突变A= 292;长度不变，但构象改= ;变，PAGE的迁移率改变

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三、= ;基因诊断的应用

诊断疾病：遗传病= 142;病、多基因疾病、恶= ;性肿瘤和感染性疾病
HLA的基因分型：

个体遗传基因的鉴= 450;：DNA指纹分析、HLA基因&= #20998;型、红细胞血型基= 240;检测、各种DNA多态性= 998;析

结构基因及其基因"= 920;达的分析

性别鉴定：性连锁$= 951;传病分析、法医学的= ;个人鉴定、运动员的&#= 24615;别检查

生物学分类鉴定

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第二= ;节基因治疗=

一、基因治疗的概= 565;

二、基因治疗的基= 412;程序

三、基因治疗的应= 992;与展望

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基因治疗的基本方法= ;

基因置换：将致病= 522;因整个地以正常基因= ;置换，使致病基因永&#= 20037;性得到更正。

基因修正：纠正致= 149;基因的突变碱基顺序= ;，而将正常部分予以&#= 20445;留。

基因修饰：将目的= 522;因导入病变细胞或其= ;他细胞，以目的基因&#= 30340;表达产物来补偿缺Ɓ= 19;细胞的功能或使原来= 的功能得到加强。

基因失活：应用反= 041;技术特异封闭某些基= ;因的表达，以抑制某&#= 20123;有害基因有表达

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二、= ;基因治疗的基本程序

（一）治疗性基因= 340;选择

（二）基因载体的$= 873;择

（三）靶细胞的选= 321;

（四）基因转移

（五）外源基因表$= 798;的筛检

（六）回输体内

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获得= ;目的基因的方法

基因的克隆化

基因的人工合成

基因的PCR扩增

人体基因组的降解

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载体= ;的选择

物理法

化学法

融合法

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受体= ;细胞（靶细胞）的种&#= 31867;

淋巴细胞

造血细胞

上皮细胞

角质细胞

内皮细胞

成纤维细胞

肝脏细胞

肿瘤细胞

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将目= ;的基因导入靶细胞

物理学方法

化学方法

融合法

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将目= ;的基因导入靶细胞

DNA磷酸钙共沉淀法

显微注射法

基因枪(粒子轰击基= 2240;转移技术)

直接注射法

电激转移

染色体介导转移法

受体介导转移法

逆转录病毒介导转= 227;法

腺病毒介导转移法

痘苗病毒介导转移= 861;

同源重组介导的靶= 521;整合(基因打靶)

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标记= ;基因技术

将目的基因和标记= 522;因重组在同一或不同= ;表达载体, 导入受体细胞, 适时&#= 36873;用选择性培养基, 筛选标记基因表型, &#= 20351;导入外源基因的靶ń= 54;胞存活,而未转导的细= ;胞则不能生存.

如G418筛选

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G418筛= 6873;原理

新霉素是细菌抗生 = 032;, 可干扰原核生物的"= 507;白质合成, 但对真核细胞则无= 433;响。

G418是新霉素的类似物= ，对原核和真核细胞= 2343;有毒性。

细菌的新霉素磷酸$= 716;移酶(neo)基因在真核细&#= 32990;中表达, 当neo基因与能有效转&= #24405;的真核DNA序列连锁时&= #23601;能获得有效有表达= 290;

产生的新霉素磷酸$= 716;移酶可使G418失活。

当真核细胞转入neo基&= #22240;后，就会含G418的选择= 性培养基中存活，而= 6410;转入neo基因的细胞则= 9981;能生存。

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三、= ;基因治疗的应用与展&#= 26395;

基因治疗的构想阶= 573;（1953～）

基因治疗的实验阶= 573;（1980 ～）

基因治疗的临床应= 992;阶段（1989.1.19 ～; 国内1991.12.31开始血友= 病的基因治疗)

基因治疗的意义和= 637;望

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人类= ;基因治疗的意义

免疫调节: 将抗原的&#= 22522;因或抗体经基因治ĭ= 03;途径导入人体,以改变= ;人体免疫反应应答和&#= 35843;节,对疾病的防治有%= 325;要意义. 将细胞因子基因导= 837;人体进行抗病毒和抗= ;肿瘤的基因治疗是目&#= 21069;基因治疗研究非常ė= 63;跃的领域。

遗传缺陷的纠正：= 532;一次为基因缺陷的遗= ;传病提供了治愈的机&#= 20250;

分泌治疗性的蛋白$= 136;，发挥治疗作用

封闭有害基因的表$= 798;：以反义技术或反义= ;失活效应抑制、封闭&#= 32959;瘤基因和病毒基因ʌ= 92;治疗病毒性和肿瘤性= 疾病。

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实现= ;基因治疗面临的困难

基因缺陷病的早期#= 786;断还很困难

基因载体本身以及= 806;源性基因整合到宿主= ;细胞染色体的位点多&#= 26159;随机的

导入的基因在宿主 = 454;胞内的表达水平难于= ;控制，表达也有一定&#= 30340;随机性

对于种类更多的、= 149;人数量更大和遗传易= ;感性疾病其基因治疗&#= 30340;难度更大

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基因= ;治疗的前景

前景光明

任重而道远

21世纪的人类基因治&#= 30103;，就像20世纪的免疫= 9044;防和抗生素的应用ᦁ= 2;将给人类的健康事业&= #24102;来极其深刻的影响

 = = = = = = = = = = = = = = = = = = = = = = = = = =
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1
2	第一= 节基因诊断= 一、基因诊断的概= 565;和特点二、基因诊断的常= 992;技术方法三、基因诊断的应= 992;
3	（一= ）基因诊断的概念: 利用现代分子生物学&= #25216;术和分子遗传学的= 216;术方法，直接检测基= ;因结构及其表达水平&#= 26159;否正常，从而对疾ĭ= 49;作出诊断的方法。一、基因诊断的概= 565;和特点
4	（二= ）基因诊断的特点= 针对性强特异性高灵敏度高适用性强
5	二、= 基因诊断的常用技术= 6041;法（一）、核酸分子= 434;交技术（二）、聚合酶链= 453;应(PCR) （三）、基因测序
6	（一= ）核酸分子杂交技术限制性内切酶酶谱= 998;析法 DNA限制性片段长度多&= #24577;性分析等位基因特异寡核!= 527;酸探针杂交法
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8	限制= 性内切酶酶谱分析法= 0030;例镰形细胞贫血原因A= 306;β-珠蛋白链第6位的GAGU= 94;GTG，造成Glu→Val所致。
9	限制= 性内切酶酶谱分析法= 0030;例镰形细胞贫血
10	限制= ;性内切酶酶谱分析法&#= 20030;例镰形细胞贫血
11
12	3、౔= 1;位基因特异寡核苷酸&= #25506;针杂交法设计针对突变和正= 120;的寡核苷酸探针分别= ;进行杂交在严格条件下进行= 434;交和洗膜
13	（二= ;）聚合酶链反应(PCR) 扩增片段长度的变= 322;：有插入或缺失时，= ;扩增长度会增加或变&#= 23567;，与正常长度比较ʌ= 92;找出致病基因 PCR-RFLP分析：相差1个碱= 2522;，改变酶切位点，ࢰ= 0;此经酶切后会产生正&= #24120;基因扩增的扩增产= 289;长度不同的片段 PCR-ASO(SSO)分析：按严格条= 件进行杂交，找出致= 0149;基因。有多种探针ࡤ= 7;进行反向斑点杂交技&= #26415; PCR-序列分析法：对于= 未知突变或少见的突= 1464;，可直接测序确定引物特异性PCR法：设&= #35745;2对引物 PCR-SSCP分析法：有突变A= 292;长度不变，但构象改= ;变，PAGE的迁移率改变
14
15	三、= ;基因诊断的应用诊断疾病：遗传病= 142;病、多基因疾病、恶= ;性肿瘤和感染性疾病 HLA的基因分型：个体遗传基因的鉴= 450;：DNA指纹分析、HLA基因&= #20998;型、红细胞血型基= 240;检测、各种DNA多态性= 998;析结构基因及其基因"= 920;达的分析性别鉴定：性连锁$= 951;传病分析、法医学的= ;个人鉴定、运动员的&#= 24615;别检查生物学分类鉴定
16	第二= ;节基因治疗= 一、基因治疗的概= 565; 二、基因治疗的基= 412;程序三、基因治疗的应= 992;与展望
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18	基因治疗的基本方法= ; 基因置换：将致病= 522;因整个地以正常基因= ;置换，使致病基因永&#= 20037;性得到更正。基因修正：纠正致= 149;基因的突变碱基顺序= ;，而将正常部分予以&#= 20445;留。基因修饰：将目的= 522;因导入病变细胞或其= ;他细胞，以目的基因&#= 30340;表达产物来补偿缺Ɓ= 19;细胞的功能或使原来= 的功能得到加强。基因失活：应用反= 041;技术特异封闭某些基= ;因的表达，以抑制某&#= 20123;有害基因有表达
19	二、= ;基因治疗的基本程序（一）治疗性基因= 340;选择（二）基因载体的$= 873;择（三）靶细胞的选= 321; （四）基因转移（五）外源基因表$= 798;的筛检（六）回输体内
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21	获得= ;目的基因的方法基因的克隆化基因的人工合成基因的PCR扩增人体基因组的降解
22
23	载体= ;的选择物理法化学法融合法
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25	受体= ;细胞（靶细胞）的种&#= 31867; 淋巴细胞造血细胞上皮细胞角质细胞内皮细胞成纤维细胞肝脏细胞肿瘤细胞
26
27	将目= ;的基因导入靶细胞物理学方法化学方法融合法
28	将目= ;的基因导入靶细胞 DNA磷酸钙共沉淀法显微注射法基因枪(粒子轰击基= 2240;转移技术) 直接注射法电激转移染色体介导转移法受体介导转移法逆转录病毒介导转= 227;法腺病毒介导转移法痘苗病毒介导转移= 861; 同源重组介导的靶= 521;整合(基因打靶)
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30	标记= ;基因技术将目的基因和标记= 522;因重组在同一或不同= ;表达载体, 导入受体细胞, 适时&#= 36873;用选择性培养基, 筛选标记基因表型, &#= 20351;导入外源基因的靶ń= 54;胞存活,而未转导的细= ;胞则不能生存. 如G418筛选
31	G418筛= 6873;原理新霉素是细菌抗生 = 032;, 可干扰原核生物的"= 507;白质合成, 但对真核细胞则无= 433;响。 G418是新霉素的类似物= ，对原核和真核细胞= 2343;有毒性。细菌的新霉素磷酸$= 716;移酶(neo)基因在真核细&#= 32990;中表达, 当neo基因与能有效转&= #24405;的真核DNA序列连锁时&= #23601;能获得有效有表达= 290; 产生的新霉素磷酸$= 716;移酶可使G418失活。当真核细胞转入neo基&= #22240;后，就会含G418的选择= 性培养基中存活，而= 6410;转入neo基因的细胞则= 9981;能生存。
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33	三、= ;基因治疗的应用与展&#= 26395; 基因治疗的构想阶= 573;（1953～）基因治疗的实验阶= 573;（1980 ～）基因治疗的临床应= 992;阶段（1989.1.19 ～; 国内1991.12.31开始血友= 病的基因治疗) 基因治疗的意义和= 637;望
34	人类= ;基因治疗的意义免疫调节: 将抗原的&#= 22522;因或抗体经基因治ĭ= 03;途径导入人体,以改变= ;人体免疫反应应答和&#= 35843;节,对疾病的防治有%= 325;要意义. 将细胞因子基因导= 837;人体进行抗病毒和抗= ;肿瘤的基因治疗是目&#= 21069;基因治疗研究非常ė= 63;跃的领域。遗传缺陷的纠正：= 532;一次为基因缺陷的遗= ;传病提供了治愈的机&#= 20250; 分泌治疗性的蛋白$= 136;，发挥治疗作用封闭有害基因的表$= 798;：以反义技术或反义= ;失活效应抑制、封闭&#= 32959;瘤基因和病毒基因ʌ= 92;治疗病毒性和肿瘤性= 疾病。
35	实现= ;基因治疗面临的困难基因缺陷病的早期#= 786;断还很困难基因载体本身以及= 806;源性基因整合到宿主= ;细胞染色体的位点多&#= 26159;随机的导入的基因在宿主 = 454;胞内的表达水平难于= ;控制，表达也有一定&#= 30340;随机性对于种类更多的、= 149;人数量更大和遗传易= ;感性疾病其基因治疗&#= 30340;难度更大
36	基因= ;治疗的前景前景光明任重而道远 21世纪的人类基因治&#= 30103;，就像20世纪的免疫= 9044;防和抗生素的应用ᦁ= 2;将给人类的健康事业&= #24102;来极其深刻的影响