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红树林土壤微生物宏基因组文库的构建及两种新颖的β-葡萄糖苷酶基因的鉴定

, PP. 168-172

Keywords: β-葡萄糖苷酶,纤维素酶,红树林,宏基因组文库

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Abstract:

宏基因组技术是挖掘微生物新酶的重要途径。通过构建红树林土壤微生物Fosmid文库,共获得了约100000个阳性克隆,该文库外源插入片段平均长度约为30kb,文库容量达3Gb。通过对文库中10000个克隆进行功能筛选,获得了17个具有β-葡萄糖苷酶活性的克隆。对其中2个表达β-葡萄糖苷酶的克隆进行亚克隆,获得2个新颖的β-葡萄糖苷酶的基因,分别命名MhGH3和bgl66。生物信息学分析表明,MhGH3基因由1992个碱基对组成,bgl66基因由2025个碱基对组成。在氨基酸水平上,MhGH3和bgl66编码的蛋白质与GenBank数据库中已知β-葡萄糖苷酶的一致性分别为64%和55%。

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