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  

Construction of a Mangrove Soil Metagenome Library and Identification of Two Novel β-Glucosidase Genes

作　　者：麦志茂[1] 苏宏飞[1] 李丽珍[1] 张偲[1] 

机构地区：[1]中国科学院南海海洋研究所中国科学院海洋生物资源可持续利用重点实验室,广州510301

出　　处：《生物技术通报》2014年第6期168-172,共5页Biotechnology Bulletin

基　　金：国家重点基础研究发展计划(“973”)(2010CB833801);国家自然科学基金重点项目(41230962)

摘　　要：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100 000个阳性克隆,该文库外源插入片段平均长度约为30 kb,文库容量达3 Gb。通过对文库中10 000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1 992个碱基对组成,bgl66基因由2 025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%。Metagenomics provides a means of exploring new enzymes in microorganisms. A mangrove soil metagenome library with 100 000 clones was constructed. The average length of inserted DNA fragment is about 30 kb, and the total capacity of inserted DNA reached about 3 Gb. Seventeen clones withβ-glucosidase activity were detected by screening 10 000 fosmid clones. The subcloning and sequencing revealed two novelβ-glucosidase genes（MhGH3 and bgl66）. Bioinformatics analysis indicated that MhGH3 and bgl66 composed of 1 992 bp and 2 025 bp in length, respectively. The deduced amino acid sequence of MhGH3 and bgl66 showed the highest sequence identity of 64%and 55%with the knownβ-glucosidase in GenBank datebase, respectively.

关 键 词：Β-葡萄糖苷酶 纤维素酶 红树林 宏基因组文库 

分 类 号：Q78[生物学—分子生物学]