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山羊瘤胃微生物宏基因组文库中一个新型纤维素酶 基因的克隆与表达
姜海琴,程建波,王力生,李吕木,范彩云
0
(安徽农业大学动物科技学院,合肥 230036)
摘要:
通过功能筛选方法,从安徽白山羊瘤胃微生物宏基因组文库中筛选得到了6个纤维素酶的阳性克隆。对其中的1个阳性克隆进一步进行亚克隆和序列分析,获得一个新型纤维素酶基因开放阅读框,通过BLAST对基因全序列进行分析比较,发现其与来自 Bacteroides cellulosilyticus DSM 14838的hypothetical protein具有51%的序列相似性和65%的同源性。并以pET28a(+)为载体、Escherichia coli BL21(DE3)为宿主菌,对新型酶基因进行高效重组表达,并对该酶表达条件进行优化。
关键词:  瘤胃微生物  宏基因组  纤维素酶
DOI:10.13610/j.cnki.1672-352x.20150302.012
投稿时间:2014-04-28
基金项目:安徽省自然科学基金项目(090411019), 安徽省教育厅自然科学基金(KJ2013A115)和高校博士点基金项目(20133418120002)共同资助。
Cloning and expression of a new cellulase gene from the metagenomic library of the goat rumen
JIANG Haiqin,CHENG Jianbo,WANG Lisheng,LI Lvmu,FAN Caiyun
(School of Animal Science and Technology, Anhui Agricultural University, Hefei 230036)
Abstract:
Six positive clones with cellulase activities were isolated from a metagenomic library of the goat rumen using a functional screening method. One positive clone was subject to further subcloning and sequence analysis and a new cellulase gene open reading frame was obtained. After blast analysis, it was found that the cellulase gene showed 51% sequence similarity and 65% homology compared with the hypothetical protein of Bacteroides cellulosilyticus DSM 14838. Using pET28a (+) as a carrier, Escherichia coli BL21 (DE3) as a host strain, the new gene was recombined and expressed efficiently and the expression conditions of the newly-cloned gene were optimized.
Key words:  rumen microbes  metagenomic  cellulase

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