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ID:36624717
大小:1.07 MB
页数:46页
时间:2019-05-13
《大鼠β防御素12的cDNA克隆及黄芪多糖对其表达调节作用的实验研究》由会员上传分享,免费在线阅读,更多相关内容在学术论文-天天文库。
1、兰州医学院硕士学位论文大鼠β-防御素-1,2的cDNA克隆及黄芪多糖对其表达调节作用的实验研究姓名:李萍申请学位级别:硕士专业:病原生物学指导教师:景涛2003.6.1≯’514853大暴昼。防锋素.1,2的cDNA亮囊及羹甍多糖对其囊选谲节作焉的实验研究摘要目的;完成昼.防御素.1,2基因的eDNA克隆,探讨黄芪多糖对大鼠呼吸系统防御素表达的调节作用。方法:跌GenBank检索到Wistar大鬣13.防御索。1,2eDNA序列,据此设计两对特异引物,采用逆转录。聚合酶链反应在Wistar大鼠气管、
2、鹏组织的总RNA中,分别扩增其特异cDNA片段,然后进行克隆扩增、限制性内切酶谱分析鞫DNA序歹
3、j测定;将6穆只健康Wistar大鼠随机分成对照组和实验组,每组30只。实验组经2.25%黄苠多糖溶液灌雹;对照组以等量生理盐水州s)灌臀,分剐乎第0夺时、第1天、第3天、第5天、第7天处死,提取其肺组织总RNA,用RT-PCR法检测大鼠B.防御素.2基因的袭达变化。结果:在大鬣气管、臃组织分别扩增斑长度为272bp,217bp的eDNA片段,其赙PCR产物经测序分析证明分别是大鼠13.防御素.1cDN
4、A与大鼠B.防御素.2cDNA。黄芪多糖实验组p.防御素.2mRNA的表达高于对照组(P5、REGULATIONOFrBD.2GENEEXPRESSIONBYASTRAGALUSPOLYSACCHARmESABSTRACTPurpose:TofulfilthecloningofratB-defensin(rBDl-1,2cDNAintrachea,lungtissueandtoexploretheregulationofrBD-2geneexpressioninlungtissuebyAstragsdusPolyr,accharides(APS).Methods:Theexperiment6、wasmadeoftwoparts.Inthefirstpart,thetotalRNAWasisolatedfromrattracheaandlung.TherBD-1,2cDNAfragmentswereobtainedbyRT-PCRamplicationwithspecificprimersrespectively.TheRT—PCRproductswereclonedinpGEM·TEasyvector.RestrictionendonucleasepaRemanalysisofthere7、combinantplasmidandDNAsequencingwereperformed.Inthesecondpart,sixtyWistarratsweighing1509wererandomlydividedintothecontrolgroupandtheAPSgroupwith30ratseach.InAPSgroup,eachwasdealtwith2.25%APSig:ThesamedoseofNSwasgiventotheratsofthecontrolgroup.Lungswer8、eharvestedimmediately,andsubsequentlyon1n,3一,5mand7mdayrespectivelyafterstimulation.TotalRNAWasextractedfromtrachealandpulmonarytissueandrBD-2wasmeasuredbyRT-PCR.口-如tinwasusedasintemalstandard.Results:TheeDNAsequenceandtheorientationoftheinsertwereconf9、irmedbyDNAsequencing.TherBD-2mRNAexpressioningroup,showedamarkedincreaseascomparedwiththatinthecontrolgroup,andalsoon3rdday,comparedwithl或,5廿land7出dayafterstimulation.Conclusions:TheseresultsindicatedthatrBD-1,rBD-2geneswereexpressedint
5、REGULATIONOFrBD.2GENEEXPRESSIONBYASTRAGALUSPOLYSACCHARmESABSTRACTPurpose:TofulfilthecloningofratB-defensin(rBDl-1,2cDNAintrachea,lungtissueandtoexploretheregulationofrBD-2geneexpressioninlungtissuebyAstragsdusPolyr,accharides(APS).Methods:Theexperiment
6、wasmadeoftwoparts.Inthefirstpart,thetotalRNAWasisolatedfromrattracheaandlung.TherBD-1,2cDNAfragmentswereobtainedbyRT-PCRamplicationwithspecificprimersrespectively.TheRT—PCRproductswereclonedinpGEM·TEasyvector.RestrictionendonucleasepaRemanalysisofthere
7、combinantplasmidandDNAsequencingwereperformed.Inthesecondpart,sixtyWistarratsweighing1509wererandomlydividedintothecontrolgroupandtheAPSgroupwith30ratseach.InAPSgroup,eachwasdealtwith2.25%APSig:ThesamedoseofNSwasgiventotheratsofthecontrolgroup.Lungswer
8、eharvestedimmediately,andsubsequentlyon1n,3一,5mand7mdayrespectivelyafterstimulation.TotalRNAWasextractedfromtrachealandpulmonarytissueandrBD-2wasmeasuredbyRT-PCR.口-如tinwasusedasintemalstandard.Results:TheeDNAsequenceandtheorientationoftheinsertwereconf
9、irmedbyDNAsequencing.TherBD-2mRNAexpressioningroup,showedamarkedincreaseascomparedwiththatinthecontrolgroup,andalsoon3rdday,comparedwithl或,5廿land7出dayafterstimulation.Conclusions:TheseresultsindicatedthatrBD-1,rBD-2geneswereexpressedint
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