Ⅱ型胶原蛋白基因cDNA序列分析及其真核表达载体的构建.pdfVIP

Ⅱ型胶原蛋白基因cDNA序列分析 及其真核表达载体的构建 王安字1，魏良纲1，乔艺杰1。张超1，张定安1。何光志2 (1贵阳中医学院第二附属医院，贵阳550002；2贵阳中医学院基础医学院) 建hCⅡ基因真核表达载体PcDNA3．1(+)／hCII，并进行PCR鉴定、酶切鉴定和序列分析。结果PCR鉴定、酶切 鉴定和序列分析表明CⅡ成功插入表达载体PcDNA3．1(+)。结论本研究成功构建了hcⅡ基因真核表达载体 PcDNA3．1(+)／hCII。 关键词：Ⅱ型胶原蛋白摹困；序列分析；真核表达载体 中图分类号：R684．3文献标志码：A 文章编号：1002_266X(2012)28瑚35．03 of cDNAand II construction Sequenceanalysistypecollagenprotein vector Of．ts’eukaryoticexpression WANG An-yul，WEl ri-j诂，ZHANGChao，ZHANGDing一矾，HEGuang-zhi Liang-gang，QIAO TheSecond (1 AffiliatedHospitalofCuiyangCollege，Guiyang550002，P．R．China) all To00nslruot vector of 1I Abstract：ObjecliveeukaryotieexpressingPeDNA3．1(+)／genetype colhscn(C1I) TotalRNAfromarticular cellsin withrheumatoidarthritiswasextractedandtheC protein．Methods cartilage patients II method．The was weD8 lit—PCR identified．andinseamJritethe genefragBleiitsamplifiedby sequencing eukaryotieexpres— sionvector constructthehumanCII vector II．And PcDNA3．1(+)to eukaryofic PcDNA3．I(+)／he gene expression constructionof vectormidentified endonucleaseand expression byPCR，restriction digestionlm[1lenceanalysis．Result C11 WaS insertedintothe vector endonudease gene successfully expressionPcDNA3．1(+)byPCR，restriction diges— tionand This has cons幻actedthe vectorPeD· sequence studysuccessfully eukaryotic analysis．Conclusion expre蟠ion NA3．I(+)／hCⅡ． of II Vectof Key typeco