Cupriavidus necator  JMP134 细菌人工染色体基因组文库的构建与鉴定

doi:10.3969/j.issn.1007-2861.2011.05.019

上海大学学报(自然科学版) ›› 2011, Vol. 17 ›› Issue (5): 675-680.doi: 10.3969/j.issn.1007-2861.2011.05.019

Cupriavidus necator JMP134 细菌人工染色体基因组文库的构建与鉴定

张俊亚，宋任涛，许政暟，朱晨光

上海大学生命科学学院上海市能源作物育种及应用重点实验室，上海 200444

收稿日期:2010-03-31 出版日期:2011-10-26 发布日期:2011-10-26
通讯作者: 朱晨光（1978~），男，副研究员，博士，研究方向为微生物功能基因鉴定分析. E-mail:cgzhu@shu.edu.cn E-mail:cgzhu@shu.edu.cn
基金资助:
上海市科委基金资助项目（08DZ2270800）；上海市高校选拔优秀青年教师科研专项基金资助项目（SHU08085）；上海大学创新基金资助项目

Construction and Characterization of Bacterial Artificial Chromosome Library of Cupriavidus necator JMP134

ZHANG Jun-ya,SONG Ren-tao,XU Zheng-kai,ZHU Chen-guang

Shanghai Key Laboratory of Bio-energy Crops, School of Life Sciences, Shanghai University, Shanghai 200444, China

Received:2010-03-31 Online:2011-10-26 Published:2011-10-26

摘要/Abstract

摘要： Cupriavidus necator JMP134(C.necator JMP134)可以降解60多种芳香族化合物，在环境污染治理方面具有良好的应用前景.该菌株基因组含有两个苯酚降解基因簇，克隆并研究其功能具有重要的理论和应用价值.以pIndigo-BAC 5为载体，构建了C.necator JMP134的细菌人工染色体(bacterial artificial chromosome,BAC)基因组文库.该文库约有9 600个克隆，插入频率为99%，平均插入片段大小达到65 kb，由此推测该BAC文库覆盖该菌株基因组约860倍.通过PCR筛选该文库的800个单克隆，得到11个含有C.necator JMP134菌株的苯酚羟化酶大亚基基因phlN2的阳性克隆.研究结果表明，该文库是一个高质量、高覆盖率的BAC文库，能有效用于目的基因簇的分离.

关键词: Cupriavidus necator JMP134, 苯酚降解, 细菌人工染色体文库

Abstract: Cupriavidus necator JMP134 (C.necator JMP134) is a model for biodegradation of aromatic compounds. The function of phenol degradation of this strain relies on a whole gene cluster. Therefore, isolation of the gene cluster is important. A bacterial artificial chromosome (BAC) library from C. necator JMP134 with vector pIndigo-BAC 5 was constructed. The library consisted of about 9 600 clones, 99% of which having an insertion with average insertion size of 65 kb. The BAC library was estimated to have 860 times of genome size. After screening one tenth of the total library clones with PCR using the designed specific primers, eleven phlN2 positive clones were obtained. The result indicates that the BAC library has high quality and high coverage, and is sufficient for isolation of target genes.

Key words: Cupriavidus necator JMP134, bacterial artificial chromosome (BAC) library, phenol biodegradation

中图分类号:

Q 78

张俊亚，宋任涛，许政暟，朱晨光. Cupriavidus necator JMP134 细菌人工染色体基因组文库的构建与鉴定[J]. 上海大学学报(自然科学版), 2011, 17(5): 675-680.

ZHANG Jun-ya,SONG Ren-tao,XU Zheng-kai,ZHU Chen-guang. Construction and Characterization of Bacterial Artificial Chromosome Library of Cupriavidus necator JMP134[J]. Journal of Shanghai University（Natural Science Edition）, 2011, 17(5): 675-680.

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Cupriavidus necator JMP134 细菌人工染色体基因组文库的构建与鉴定

Construction and Characterization of Bacterial Artificial Chromosome Library of Cupriavidus necator JMP134

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