荷斯坦奶牛瘤胃微生物元基因组Fosmid文库的构建与分析

Construction and Analysis of Genomic Fosmid Library from Rumen Microbes in Holstein Cow

采用包埋法提取荷斯坦奶牛瘤胃微生物大片段总DNA,纯化后脉冲场电泳回收大小为36-48 kb,与pcc2FOS vector连接,转染至大肠埃希菌EPI 300宿主细胞,构建瘤胃微生物Fosmid基因组文库。对文库进行鉴定,该文库平均插入片段大小约35 kb,共保存30 000个克隆,空载体率小于2%,库容达1050 Mb。

Total large fragment genomic DNA extracted by embedding method from rumen microbes was prepared for library construction after purification 36-48 kb of was recovered by pulsed field electrophoresis and ligated with pcc2FOS vector and transformed into E.coli EPI300.A Fosmid genomic library of rumen microbe was successfully constructed with the capacity of 1 050 Mb in which the average inserted fragment size at about 35 kb,and totally stored 30 000 clones with the empty rate at lower than 2%.