鹿茸间充质细胞长cdna文库的构建.pdf

摘 要 本试验以梅花鹿二杠茸为研究材料，通过组织学解剖，取得鹿茸的间充质， 经过胰酶消化，培养鹿茸间充质细胞，成功构建了鹿茸间充质细胞的cDNA文库。 物经体外蛋白包装，产生初级文库；鉴定文库的滴度后，进行文库扩增。随机挑 取若干个噬菌斑，进行PCR扩增，检测所构建的cDNA文库的质量。经测定该文 库初始滴度为3．O×l 得到反转录，得到的cDNA是完整的，完全符合eDNA文库的要求。 鹿茸间充质细胞全长eDNA文库的成功构建，为我们钓取鹿茸间充质特异基 因研究奠定了坚实的基础，同时对鹿茸的再生机理方面也会有所帮助。 关键词 鹿茸，问充质，细胞培养，cDNA文库 Abstract This obtainedthe fromtheantlerthe experiment mesenchyme by histology theantler dissection．Thenwe thematerialwiththe andcultured digested trypsin series toconstructthe takeda of mesenchymalcells．Subsequently，weexperiments eDNA thetotalRNAformantler cellswith library，suchas，extracting mesenchymal to stranded doublestrand TRIZOL，reversetranscriptiongetsingle eDNA，synthesis withLD—PCR PCR afterthe K eDNA method；purificationproduct proteinase withtheSill the hydrolysis，cuttingenzyme；classificationsepearationdigested theeDNA above andconnectionwith product，retrievingcomponent500bp XTriplEx2 connectionafterinvitro the primary carrier；The product proteinpacking，producing the the to library；After titer，takinglibraryamplification． identificatinglibrary a numberof theuniversal of Randomlychoosing macrophagesplaque，with