摘要
采用宏基因组技术构建了高糖土壤微生物的DNA文库，该文库约含9万个克隆，文库外源DNA总容量为3．1×10%^9bp。利用活性筛选策略，对文库进行筛选，获得11个β-葡萄糖苷酶的阳性克隆，并对其中2个表达β-葡萄糖苷酶的克隆进行亚克隆和序列分析，获得两个编码新型B一葡萄糖苷酶的基因分别命名为：Mn6印引和Mn6印3日。生物信息学分析表明：“unbgl3A基因由2241个碱基对组成，unbgl3B基因由2292个碱基对组成。在核苷酸水平上，unbgt3A、unbgl3B与已知数据库中的β-葡萄糖苷酶基因没有任何相似性。在氨基酸水平上，与GenBank数据库中已知β-葡萄糖苷酶的相似性分别为73％~1169％。
A metagenomic DNA library of microbes from sugar enriching soil was constructed, which contained 90 000 clones; the capacity of inserted DNA in thislibrary reached about 3.1 x 10^9 bp. Eleven clones expressing [β-glucosidase activity were isolated by function-based screening strategy and two clones of them were subcloned and analyzed in sequence. The results revealed that there were two novel β-glucosidase genes, named as unbgl3A and unb gl3B Bioinformatics analysis indicated that unbgl3A should be composed of 2 241 base pairs in length, while unbgl3B should be composed of 2 292 base pairs in length. There is no homologous sequences between unbgl3A or unbgl3B compared to with the sequences of the β-glucosidase genes in GenBank database, whereas unbgl3A and unbgl3B showed 73% and 69% identities with the known protein sequences in GenBanlc
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2013年第1期30-35,共6页
Genomics and Applied Biology
基金
广西自然科学基金“利用木糖生产燃料酒精的基因工程菌构建及木糖代谢途径研究”(2010GXNS-FA013067)资助
