用λgt11载体构建人脑胶质瘤基因文库  

Construction of the Human Glioma cDNA Library Using λgt11 Vector

作　　者：王尧[1] 覃文新[1] 左剑玲[1] 韦永新 陈桂林[1] 李斌[1] 杜子威[1] 赵涵芳[2] 章有章[2] 程枫[2] 陈诗书[2] 

机构地区：[1]苏州医学院脑神经研究室 [2]上海第二医科大学生物化学教研室分子生物学实验室

出　　处：《苏州医学院学报》1992年第3期179-182,254-255,共4页Acta Academiae Medicinae Suzhou

基　　金：国家自然科学基金

摘　　要：自人脑胶质瘤手术标本中提取mRNA,逆转录合成cDNA,在T_4DNA 连接酶作用下,与一头是平末端,另一头是EcoR I 粘性末端、内含Not I 切点的插头相连,经T_4多聚核苷酸激酶磷酸化后,再与脱磷酸处理过的λgt11 臂连接,形成重组λgt11DNA。体外包装,构建了一个库容量含1.02×10~6重组子的人脑胶质瘤cDNA 基因文库。克隆效率为1.08×10~7pfu/μg cDNA。重组百分比为96.2%。RNA were purified from two specimens of human glioma obtained from operation.The double—stranded cDNA molecules were synthesized by reverse transcription.The eDNA molecules were lig-ated to adaptors containing an internal Not I site,a blunt end and a preformed EcoR I cohesive endat the other end.After the adapted cDNA molecules had een kinased,it was able to be ligated to dephosphorylated λ gtll vector arms.This resulted in recombinant λgtll DNA molecules.The hymanglioma cDNA library was constructed by in vitro packaging.The library contained 1.02×10~5 reco-mbinants.Cloning efficiency was 1.08×10~7 pfu/μg cDNA.Percentage of recombinants was 96.2%.

关 键 词：CDNA 基因文库 脑肿瘤 胶质瘤 

分 类 号：R739.410.3[医药卫生—肿瘤]