茁霉多糖酶基因的克隆  

Cloning Of K. aerogenes Pullulanase Gene

作　　者：何玲[1] 罗进贤[2] 

机构地区：[1]广州医学院化学致癌研究所 [2]中山大学生物工程中心

出　　处：《广州医学院学报》1994年第5期1-4,共4页Academic Journal of Guangzhou Medical College

摘　　要：以λEMBL3DNA为载体，产气克氏杆菌染色体的部分酶切片段为供体，成功地构建了产气克氏杆菌的基因文库，重组噬菌体数为1．7×103ρfu／ugDNA，达到了Clarke与Carbon公式的要求。经支链淀粉平板初筛和茁霉多糖平板复筛，从此文库中得到三个产茁霉多糖酶的噬菌体。酶切分析证明这三个重组噬菌体含有相同的酶切片段。由此我们构建了产气克氏杆菌的基因文库，并从中钓到了茁霉多糖酶基因。A complete genomic library of K.aerogenes was successfully construeted using λ EMBL3 DNA as vector and K.aerogenes chromsome DNA as donor. The cloning effeciency was 1.7 × 103 pfu/μg DNA, which reached the number of clones reqired by Clarke and Carbon formula. Three pullulanase positive recombinant phages were obtained from this library using the amylo pectin and pullulan plate.These three phages contained the same DNA by enzymatical analysis.

关 键 词：基因文库 产气克氏杆菌 茁霉多糖酶基因 λEMBL3DNA 

分 类 号：R341[医药卫生—基础医学]