霍乱弧菌染色体基因文库的构建

CONSTRUCTION OF GENOMIC LIBRARY OF VIBRIO CHOLERAE 178

利用限制性内切酶Ｓａｎ３Ａ将霍乱弧菌１７８（埃尔托生物型，小川血清型）染色体ＤＮＡ进行部分酶切后，分离２０～５０ｋｂ的ＤＮＡ片断，与经过ＢａｍＨＩ酶切的柯斯质粒ｐＨＣ７９连接重组，通过体外包装系统形成噬菌体颗粒，感染受体菌Ｅ．ｃｏｌｉＨＢ１０１，构建成霍乱弧菌１７８染色体基因文库。为霍乱弧菌中未知基因的克隆及其它研究打下了基础。

Genomic fragments ranging from 20～50 kb were obtained by controled partial digesting thechromosomal DNA of the VI'brio cholerae 178 strain(Eltor biotype,Ogawa serotype)with restrictionendonuclease San3A.The cosmid vector PHC79 was digested with BamHI.The two DNA fragmentwere used to ligated and packaged using in vitro pa ckaging system.The packaged phage particles wereused to infect E.colt HB101 for constructing a genomic library of the V.cholerae 178.It will be beneficial for molecular cloning other unknown genes of V.cholerae.