北京红篓菌聚羟基烷酸合成酶基因的克隆与表达  

Cloning and Expression of Poly-hydroxyalkanoates Synthase Genes of Rhodocista pekingensis

作　　者：张伟[1] 姜成英[1] 戴欣[1] 刘双江[1] 

机构地区：[1]中国科学院微生物研究所,北京100080

出　　处：《微生物学报》2004年第3期332-335,共4页Acta Microbiologica Sinica

基　　金：中国科学院院长青年创新基金~~

摘　　要：以北京红篓菌基因组DNA为供体 ,pKC5 0 5cosmid质粒为载体系统 ,λ 噬菌体体外包装重组质粒并转染感受态细胞E .coliJM1 0 9,构建了该菌株基因组DNA文库。以聚羟基烷酸 (PHA)合成酶基因通用简并引物扩增的PHA合成酶部分基因为探针 ,经DIG标记后利用菌落原位杂交技术对文库进行筛选 ,获得 3个阳性克隆菌株 ,PCR检测证明这 3个克隆具有PHA合成酶基因片段 ,酶活测定表明Genomic DNA of Rhodocista pekingensis was partial digested, ligated to cosmid pKC505,packaged with λ-coat protein using a vitro packaging kit, and transfected into E.coli JM109. Using the DIG-labled probe obtained from part of the PHA synthase gene of Rhodocista pekingensis by PCR. The genomic library was screened by in situ-colony hybridization and 3 positive hybrid strains were obtained. PCR amplification and enzyme activity measurement revealed that all the 3 strains had a fragment of PHA synthase gene and the activities of PHAs synthase, NADPH-dependent acetoacetyl-CoA reductase related to synthesis of PHAs.

关 键 词：北京红篓菌 聚羟基烷酸 PHA合成酶 

分 类 号：Q786[生物学—分子生物学]