木聚糖酶产生菌的筛选鉴定及木聚糖酶基因(Xyn B)的克隆  

Screening and identification of xylanase producing strain and cloning of its Xyn B gene

作　　者：莫毅 梁方方[2] 赖志强[2] 卢玉发[2] 廖玉英[2] 何仁春[2] 黄丽霞[2] 杨琳[3] 

机构地区：[1]广西人口和计划生育研究中心 [2]广西畜牧研究所 [3]华南农业大学动物科学学院

出　　处：《中国饲料》2011年第23期20-22,共3页China Feed

基　　金：广西青年基金项目(桂科青0991068)资助

摘　　要：采用富集培养、透明圈平板筛选的方法,从长期堆放青贮饲料的土壤中分离到7株木聚糖酶产生菌。选取透明圈最大的木聚糖酶产生菌X7作为基础菌,对其进行16S rRNA部分序列PCR扩增测定,经BLAST同源性分析确定X7为枯草芽孢杆菌Bacillus subtilis strain GD3b(Genebank编码:HM055602.1)。然后设计特异引物,以X7基因组为模板,对其进行Xyn B基因序列PCR扩增,扩增产物经BLAST同源性分析表明已成功克隆了X7菌株的Xyn B基因,为日后构建木聚糖酶基因酵母表达载体,培育能够分泌表达具有高活性的木聚糖酶酵母工程菌奠定了基础。In this study,7 xylanase producing strains were screened from the soil where silage had been piled for a long time by enriching culture,hydrolysis ring method.Then the most high-yield xylanase producing strain X7 was selected as the basic bacteria to perform the subsequent experiments： （1）To determine differentia,X7＇s 16S rRNA partial sequence was amplified by PCR method and identified by BLAST homology analysis; （2） To clone X7＂s xylanase B Xyn B gene, XT＇s genome was used as the template and two specific primers of Xyn B gene were designed to perform PCR amplifica- tion.After that, the PCR products were purified, sequenced and identified by BLAST homology analysis.Results showed that X7 was the most similar to Bacillus subtilis strain GD3b（Genebank code ：HM055602.1 ）and X7 ＇s Xyn B gene was cloned successfully.All these results laid a foundation for constructing the Pichia expression vector for Xyn B and breeding the high effective excretion recombined gene Pichia.

关 键 词：木聚糖酶 菌株筛选 16s RRNA 基因克隆 

分 类 号：S816.7[农业科学—饲料科学]