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EcoRⅠ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8.2kb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25kb,最大约为50kb,平均大小估算为40kb,文库克隆容量约52Mb,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。
The 1 329-clones genomic library of Klebsiella oxytoca KO108 was constructed by pWEBTM 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.
出处
《广西师范大学学报（自然科学版）》
CAS
北大核心
2016年第2期151-157,共7页
Journal of Guangxi Normal University:Natural Science Edition
基金
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)
