嗜热菌Aquifex pyrophilus中mbh2基因簇的鉴定和部分基因的克隆及测序  

IDENTIFICATION AND CLONING OF PARTIAL MBH2 GENE CLUSTER OFHYPERTHERMOPHILE AQUIFEX PYROPHILUS

作　　者：吕健[1] G Rakhely K L Kovacs 肖昌松[1] 周培瑾[1] 

机构地区：[1]中国科学院微生物研究所,北京100080 [2]匈牙利科学院生物研究中心生物物理研究所

出　　处：《微生物学报》2001年第6期674-679,共6页Acta Microbiologica Sinica

摘　　要：基于嗜热菌Aquifexaeolicus的mbhS2基因 ,设计合成特异性引物 ,以Aquifexpyrophilus的染色体DNA为模板 ,应用PCR技术扩增出目的片段。序列测定结果显示 ,其推导出的氨基酸序列与A .aeolicus的相应序列具有 85 %的同源性。以此PCR片段为探针 ,从A .pyrophilus的NcoⅠ部分基因组文库 (4～ 6kb)中筛选出含 5kb大小插入片段的阳性克隆 ,进而对其进行了亚克隆及测序。结果表明 ,该插入子包含A .pyrophilusmbh2基因簇的小亚基基因mbhS2、orf1基因和部分orf2基因。所推导出的氨基酸序列与A .aeolicus的MbhS2和Orf96 3相比较的同源性分别为 81 %和 6 0 %。A 0.8kb fragment of mbhS2 gene of Aquifex pyrophilus was obtained by PCR with designed primers basing mbhS2 gene of A.aeolicus.It showed 85% homology with the corresponding region of A.aeolicus. Using it as probe, a 5.0kb NcoⅠ fragment was fished out from the partial genomic library of A.pyrophilus. Then this fragment was cloned,subcloned and sequenced.The result revealed that the fragment contains the full length gene for the mbhS2,the gene orf1 and the first 366 bp of orf2. Comparison with mbhS2 and orf963 of A.aeolicus shows 81% and 60% homologies in amino acid sequence,respectively.

关 键 词：mbh2基因族 基因组文库 亚克隆 测序 鉴定 嗜热菌 

分 类 号：Q785[生物学—分子生物学]