|本期目录/Table of Contents|

[1]官兴颖,吴振芳,吴琦,等.枯草芽孢杆菌C-36内切葡聚糖酶基因的克隆及其在大肠杆菌中融合表达[J].生物加工过程,2009,7(03):68-72.
 GUAN Xing-ying,WU Zhen-fang,WU Qi,et al.Cloning,sequencing,and expression of endoglucanase gene from Bacillus subtilis C-36 in Escherichia coil[J].Chinese Journal of Bioprocess Engineering,2009,7(03):68-72.
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枯草芽孢杆菌C-36内切葡聚糖酶基因的克隆及其在大肠杆菌中融合表达(/HTML)
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《生物加工过程》[ISSN:1672-3678/CN:32-1706/Q]

卷:
7
期数:
2009年03期
页码:
68-72
栏目:
出版日期:
2009-05-30

文章信息/Info

Title:
Cloning,sequencing,and expression of endoglucanase gene from Bacillus subtilis C-36 in Escherichia coil
文章编号:
1672-3678(2009)03-0068-05
作者:
官兴颖吴振芳吴琦胥兵陈惠
四川农业大学 生命科学与理学院,雅安 625014
Author(s):
GUAN Xing-yingWU Zhen-fangWU QiXU BingCHEN Hui
College of Life Science,Sichuan Agricultural University,Ya′an 625014,China
关键词:
枯草芽孢杆菌克隆内切葡聚糖酶基因表达序列
分类号:
Q781;Q785
文献标志码:
A
摘要:
以自行分离筛选出的天然枯草芽孢杆菌(Bacillus subtilis) C-36的染色体DNA为模板,PCR扩增得到含有内切葡聚糖酶基因的DNA片段,将其克隆到pMD-18T载体中,序列分析表明,克隆得到的DNA片段全长1 602 bp,编码一个含有499个氨基酸的多肽。与其他芽孢杆菌内切葡聚糖酶基因序列比对,其核苷酸同源率为90%~93%,其编码的氨基酸序列的同源性在90%~98%,已将此基因注册GenBank(DQ782954)。将含内切葡聚糖酶基因的重组克隆质粒进行亚克隆,用KpnⅠ和EcoRⅠ双酶切后,与相同酶切的表达载体pET-32a相连接,并导入大肠杆菌BL21中表达。蛋白质电泳实验结果表明在6.47×104处有表达蛋白带。经测定表达蛋白比酶活力达99.02 U/mL,为出发菌C-36(63.78 U/mL)的1.55倍。

参考文献/References:

[1]Klyosov A A. Trends in biochemistry and enzymology of cellulose degradation[J]. Biochemistry,1990,29(47): 10577-10585.
[2]Sleat R,Mah R A,Robinson R. Isolation and characterization of an anaerobic,cellulolytic bacterium,Clostridium cellulovorans sp.nov[J]. Appl Environ Microbiol,1984,48(1): 88-93.
[3]Ponpium P,Ratanakhanokchai K,Kyu K L. Isolation and properties of a cellulosome-type multienzyme complex of the thermophilic Bacteroides sp.strain P-1[J]. Enzyme Microb Technol,2000,26(5/6): 459-465.
[4]陈兵,林开江.一株芽孢杆菌产生的碱性纤维素酶的研究[J].浙江农业学报,1994,6(1): 37-40.
Chen Bing,Lin Kaijang. Studies on alkaline cellulase produced by Bacillus sp. No.1[J]. Acta Agriculturae Zhejiangensis,1994,6(1): 37-40.
[5]沈雪亮,夏黎明,刘景晶.纤维素酶E5基因在E.coli中的克隆与表达[J].浙江大学学报:工程版,2001,35(6): 640-644.
Shen Xueliang,Xia Liming,Liu Jingjing. Recombination and expression of cellulase E5 gene[J]. Journal of Zhejiang University: Engineering Science Edition,2001,35(6): 640-644.
[6]汪维云,朱金华,吴守一.纤维素科学及纤维素酶的研究进展[J].江苏理工大学学报,1998,19(3): 20-28.
Wang Weiyun,Zhu Jinhua,Wu Shouyi. Research progress of cellulose science and cellulase[J]. Journal of Jiangsu University of Science and Technology,1998,19(3): 20-28.
[7]Sambrook J,Fritsch E F,Maniatis T. Molecular cloning a laboratory manual[M]. 2nd ed. Beijing: Science Press,1996.
[8]王晓芳,徐旭士,吴敏,等.一株纤维素分解菌的分离与筛选[J].生物技术,2001,11(2): 27-30.
Wang Xiaofang,Xu Xushi,Wu Min,et al. Isolation and screening of cellulose-decomposing microorganisms[J]. Biotechnology,2001,11(2): 27-30.
[9]Borriss R,Buettner K,Maentsaelae P. Structure of the beta-1,3-1,4-glucanase gene of Bacillus macerans: homologies to other beta-glucanases[J]. Mol Gen Genet,1990,222(2/3): 278-283.
[10]胥兵,陈惠,韩学易,等.枯草芽孢杆菌C-36纤维素酶的纯化及酶学性质[J].四川农业大学学报,2006,24(4): 398-401.
Xu Bing,Chen Hui,Han Xueyi,et al. Purification and properties of cellulase from Bacillus subtilis C-36[J]. Journal of Sichuan Agricultural University,2006,24(4): 398-401.

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备注/Memo

备注/Memo:
收稿日期:2008-09-10
基金项目:四川省科技厅应用基础研究资助项目(05JY0290361);四川省教育厅自然科学科研项目(2005A030)
作者简介:官兴颖(1982—),女,重庆人,硕士研究生,研究方向:分子酶学;陈惠(联系人),教授,E-mail:chenhui@sicau.edu.cn
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