巨大芽孢杆菌内切葡聚糖酶编码基因的克隆及序列分析

巨大芽孢杆菌内切葡聚糖酶编码基因的克隆及序列分析

黄玉杰^1,2;杨合同^1,3;丁爱云²;李纪顺¹;周红姿¹;王萍⁴;Maarten Ryder⁵

1. 山东省科学院生物研究所,济南　250014;2. 山东农业大学植物病理系;3. 山东理工大学生命科学学院;4. 山东中医药大学;5. Australia Commonwealth Scientific and Industrial Researth Organization Land and Water

收稿日期:2004-04-26 修回日期:1900-01-01 出版日期:2004-11-08 发布日期:2004-11-08

Cloning and Sequencing of Endoglucanase Gene from Bacillus megaterium

HUANG Yu-jie;YANG He-tong;DINGAi-yun;LI Ji-shun;ZHOU Hong-zi;WANG Ping;Maarten Ryder

Biology Research Institute , Shandong Academy of Sciences , Jinan 250014 , China

Received:2004-04-26 Revised:1900-01-01 Online:2004-11-08 Published:2004-11-08

摘要/Abstract

摘要： 利用pBluescripts构建巨大芽孢杆菌基因组文库,在ABP平板上测定酶活性,共有78株阳性克隆子具有内切葡聚糖酶活性。将酶活性表达较好的一株菌株进行序列测定。测定结果这个片段共941bp,含有一个开放阅读框架,共348个核苷酸,可编码116个氨基酸。序列比较结果表明,该基因片段同已发表的枯草芽孢杆菌glyB-aprE之间的同源性为35%;同芽孢杆菌BP23 celB、短小芽孢杆菌内切葡聚糖酶和多粘芽孢杆菌β-1,4-内切葡聚糖酶的编码基因的同源性只有27%。虽然同源性较低,但酶活性表达较强,认为该基因是编码内切葡聚糖酶的一个新基因片段。

Abstract: Genomic DNA of Bacillus megaterium was partially digested by restriction enzyme Sau3A and was used to establish the genomic library in plasmid pBluescripts. Seventy-four clones that showed hydrolysis ability on the ABP plate were obtained. The gene fragment of 941bp for the endoglucanase was
cloned. The fragment contains an open reading frame 348 nucleotides starting with the initiation codon ATG at position 571 and ending with the termination codon TAA at position 919 , and deduced amino acid is about 116. Sequencing analysis showed that the sequence of endoglucanase fragment exhibits 35 %
homology with B . subtilis chromosomal DNA(from glyB to aprE) , and 27 % homology with Bacillus sp. BP23 celB genes , B . pumillus endoglucanase and B . polymyxa beta-1 , 4-endoglucanase genes , respectively. It was recognized as a new gene sequence encoding for endoglucanase of B . megaterium.

黄玉杰1,2 ;杨合同1,3 ;丁爱云2 ;李纪顺1 ;周红姿1 ;王萍4 ;Maarten Ryder5 . 巨大芽孢杆菌内切葡聚糖酶编码基因的克隆及序列分析[J]. , DOI: .

HUANG Yu-jie;YANG He-tong;DINGAi-yun;LI Ji-shun;ZHOU Hong-zi;WANG Ping;Maarten Ryder. Cloning and Sequencing of Endoglucanase Gene from Bacillus megaterium[J]. , DOI: .