云南大额牛瘤胃宏基因组Fosmid文库的构建与分析

Abstract

Abstract: To study cellulose genes of Yunnan Gayal rumen,the high molecular weight DNA was extracted and purified from rumen contents of 4 adult Gayals by embedding method and pulsed field gel electrophoresis approaches. After digestion with AluⅠ, DNA fragments ranging from 23~48 kb were collected and ligated with pcc2 FOS vector and transformed into E.coli EPI 300.A rumen metagenomic Fosmid library with the capacity of 1363 Mb was successfully constructed. This library contained 38400 clones with an average inserting size of 35.5 kb,and empty rate was lower than 2%.The screening of the partial metagenomic library identified 95 clones expressing activity of carboxymethyl cellulase,52 clones with xylanase activity, and 23 clones with carboxymethyl cellulase and xylanase activity. These results provided the foundation to further study cellulase gene in Gayal rumen.

Key words: Gayal; rumen microbes; Fosmid library; screen

CLC Number:

S823

LI Bi-feng, ZHU Ya-xin, GOU Xiao, LENG Jing, MAO Hua-ming, LI Qing, FENG Li, YANG Shu-li. Construction and Analysis of Metagenomic Fosmid Library from Rumen in Yunnan Gayal[J]. , 2013, 40(12): 61-65.

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Construction and Analysis of Metagenomic Fosmid Library from Rumen in Yunnan Gayal

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