福氏2a志贺菌301株染色质基因文库的构建

目的　构建福氏 2a志贺氏菌 3 0 1株染色质DNA基因文库 ,为下一步全基因组序列测定打下基础。方法　Murry法提取福氏 2a志贺氏菌染色质DNA ,采用鸟枪法策略 ,分别经 8种限制性内切酶消化后克隆至载体质粒中 ,并经双脱氧链末端终止法测序后计算重组质粒重复率。结果　Murry法提取福氏 2a志贺氏菌染色质DNA大小为 3 0Kb以上 ,共构建 86 3 2个重组质粒 ,质粒重复率为 1 %。结论　本研究在国际上首次成功构建了福氏 2a志贺氏菌株染色质DNA基因文库 ,文库全长为福氏 2a志贺氏染色质的 6倍 ,已足够全基因组序列测定所需。

The construction of clone library of Shegillier Flexneri 2 a chromosome DNA

Objective The genome library of Shegillier Flexneri 2a chromosome DNA was constructed to fertilize the coming sequencing of the whole genome. Methods The genome library of Shigella Flexneri 2a was constructed by shotgun strategy.The chromosome genome was digested by 8 restriction endonuclease respectively,and the overlapping DNA fragments were cloned into carrying plasmid and then the rate of same recombine plasmid was counted after sequencing a set of the clones.Results The length of Shigella Flexneri 2a chromosome DNA prepared by Murry's method was more than 30?Kb. There were 8632 clones and 1% repeat clones was found.Conclusion It's the first time to construct the different overlapping gene libraries in the world.And it is enough for sequencing the whole genome.