多能硫杆菌基因文库的构建以及Rubis CO基因的筛选  被引量:1

CONSTRUCTION OF THIOBACILLUS VERSUTUS GENE LIBRARY AND ISOLATION OF RECOMBINANT CONTAINING RIBULOSE 1,5-BISPHOSPHATE CARBOOXYLASE/OXYGENASE

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作  者:刘振盈[1] 颜望明[1] 徐海岩 郑雷 

机构地区:[1]山东大学生命科学院微生物技术国家重点实验室,济南250100

出  处:《微生物学通报》1997年第2期99-103,共5页Microbiology China

基  金:国家及山东省自然科学基金

摘  要:以pUC9质粒DNA作为载体,用PstⅠ酶切、碱性磷酸酯酶处理后,与PstⅠ部分酶切的多能硫杆菌染色体DNA3-10kbDNA片段连接,转化大肠杆菌JM83,在MacConkey培养基上筛选重组于,所得的重组子为6.5×103,达到建库要求的理论值。进一步用光合细菌Rhodobactersphaeroides类型Ⅱ磷酸核酮糖羧化酶/加氧酶基因(rbcL-rbcS)为探针,从该库中筛选到了含有多能硫杆菌的1,5-二磷酸核酮糖羧化酶/加氧酶(RubisCO)基因片段的重组子,从而进一步证明所构建多能硫杆菌基因文库的完整性和可靠性。The genomic library of Thboacdha mptus was constructed by using pUC9 plasmid as vecter. pUC9 was digested by PstI enzyme, treated with phosphatase, then ligated with 3~10kb fragments of T. versutus partially digested by PstI and transformed to hota coli JM83. The recombinant were screened on MacConkey medium and 6.5 ×103 recombinants were obtained. It has exceeded the necessary number of recombinants that represented an entire genom of T.versutus.Furthermore the recombinant containing the gene fragment of ribulose 1, 5-bisphosphate carbooxylase/oxygenase was isolated from the library.

关 键 词:多能硫杆菌 基因文库 RUBISCO基因 

分 类 号:Q939.145[生物学—微生物学]

 

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