产酸克雷伯氏菌基因组文库的构建及鉴定

doi:10.16088/j.issn.1001-6600.2016.02.023

广西师范大学学报（自然科学版） ›› 2016, Vol. 34 ›› Issue (2): 151-157.doi: 10.16088/j.issn.1001-6600.2016.02.023

• • 上一篇

产酸克雷伯氏菌基因组文库的构建及鉴定

江绍锋^1,2,3, 黄金清^1,2, 于晓宇^1,2, 李云飞^1,2, 陆祖军^1,2

1.广西师范大学生命科学学院,广西桂林541006;
2.珍稀濒危动植物生态与环境保护教育部重点实验室,广西桂林541006;
3.华南农业大学农学院,广东广州510462

收稿日期:2015-07-18 出版日期:2016-06-25 发布日期:2018-09-14
通讯作者: 陆祖军(1963—),男(壮族),广西百色人,广西师范大学教授,博士。E-mail:luzujun2002@aliyun.com
基金资助:
国家自然科学基金资助项目(31060120);广西自然科学基金资助项目(2013GXNSFAA019059);广西研究生教育创新计划项目基金资助(YCSZ2014090)

Construction and Characterization of the GenomicLibrary of Klebsiella oxytoca

JIANG Shaofeng^1,2,3, HUANG Jinqing^1,2, YU Xiaoyu^1,2, LI Yunfei^1,2, LU Zujun^1,2

1.College of Life Science, Guangxi Normal University, Guilin Guangxi 541006,China;
2.Key Laboratory of Ecologyof Rare and Endangered Species and Environmental Protection(Ministry of Education),Guilin Guangxi 541006,China;
3. College of Agriculture,South China Agriculture University,Guangzhou Guangdong 510462,China

Received:2015-07-18 Online:2016-06-25 Published:2018-09-14

摘要/Abstract

摘要： 为了给基因功能研究提供基础,以pWEB^TM质粒构建产酸克雷伯氏菌KO108基因组文库,克隆数目为1 329个。通过EcoR Ⅰ酶切分析随机挑取的20个文库克隆的重组质粒,结果显示所有质粒均有8.2 kb载体条带同时含有外源DNA,且外源DNA的带型并不相同,这表明所构建的KO108基因组文库中插入的外源DNA随机性较好。分析外源DNA电泳条带的大小,结果显示克隆的外源DNA片段最小约为25 kb,最大约为50 kb,平均大小估算为40 kb,文库克隆容量约52 Mb,证明该基因组文库包含基因组任意一个基因的概率为99.8%,是KO108基因组覆盖率的5倍。该文库的构建为产酸克雷伯氏菌功能基因克隆和比较基因组学研究奠定了基础。

关键词: 产酸克雷伯氏菌, 基因组文库, pWEB^TM质粒, 覆盖率, 库容

Abstract: The 1 329-clones genomic library of Klebsiella oxytoca KO108 was constructed by pWEB^TM Cosmid. Its coverage and randomness was confirmed by the electrophoresis profiles of 20 recombination plasmids digestion with EcoR Ⅰ,which were randomly selected from the library. The results showed that all tested plasmids contained the 8.2 kb cosmid fragment and an exogenous one diverse in size from 25 to 50 kb(about 40 kb in average), this suggested that the total capacity of this library was about 52 Mb, 5 times of KO108 genome, 99.8% theoretical possibility including any gene in this strain. The library lays a foundation for functional gene cloning and comparative genomics research for Klebsiella oxytoca.

Key words: Klebsiella oxytoca, genomic library, pWEB^TM Cosmid, coverage, library capacity

中图分类号:

Q933

江绍锋, 黄金清, 于晓宇, 李云飞, 陆祖军. 产酸克雷伯氏菌基因组文库的构建及鉴定[J]. 广西师范大学学报（自然科学版）, 2016, 34(2): 151-157.

JIANG Shaofeng , HUANG Jinqing , YU Xiaoyu , LI Yunfei , LU Zujun. Construction and Characterization of the GenomicLibrary of Klebsiella oxytoca[J]. Journal of Guangxi Normal University(Natural Science Edition), 2016, 34(2): 151-157.

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产酸克雷伯氏菌基因组文库的构建及鉴定

Construction and Characterization of the GenomicLibrary of Klebsiella oxytoca

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