全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...

苏云金杆菌cry8Ea2蛋白的分离纯化及生物活性测定

DOI: 10.3969/j.issn.1000-7091.2012.06.011, PP. 48-52

Keywords: 苏云金杆菌,cry8,穿梭载体,蛋白纯化,生物活性测定

Full-Text   Cite this paper   Add to My Lib

Abstract:

cry8Ea2蛋白是一种新型苏云金杆菌杀虫晶体蛋白。通过穿梭载体pSXY422b将cry8Ea2导入Bt无晶体突变菌株中,获得工程菌HD8Ea2。cry8Ea2蛋白在工程菌中获得表达。通过分子筛层析,成功获得纯化的130kDa活性毒素蛋白。用改进的生物活性测定方法进行cry8Ea2蛋白对华北大黑鳃金龟和暗黑鳃金龟一龄幼虫的杀虫活性测定,LC50分别为0.2679μg/mL和0.0719μg/mL。研究首次发现cry8Ea2蛋白对华北大黑鳃金龟具有较高毒力,拓展了该蛋白的杀虫谱,为转基因工程提供了新资源。

References

[1]  Chattopadhyay A,Bhatnagar N B,Bhatnagar R. Bacterial insecticidal toxins[J]. Crit Rev Miicrobiol, 2004, 30( 1) : 33-54.
[2]  Fernando H V,Marliton R B. Bacillus thuringiensis survey in Brazil: geographical distribution and insecticidal activity against spodoptera frugiperda[J]. Neotropical Entomology, 2003, 32( 4) : 639-644.
[3]  Roh J Y,Choi J Y,Li M S, et al. Bacillus thuringiensisas a specific,safe,and effective tool for insect pest control[J]. Microbiol Biotechnol, 2007,7 ( 4) : 547-559.
[4]  Lambert B,Peferoen M. Insecticidal promise of Bacillus thuringiensis Facts and mysteries about a successful biopesticide[J]. BioScience, 1992, 42: 112-122.
[5]  Sharpe E S. Toxicity of the parasporal crystal of Bacillus thuringiensis to Japanese beetle larvae [J]. Invertebr Pathol, 1976, 27, 421-422.
[6]  Ohba M, Iwahana H,Asano S, et al. A unique isolate of Bacillus thuringiensis serovar japonensis with a high larvicidal activity specific for scarabaeid beetles[J]. Lett Appl Microbiol, 1992, 14, 54-57.
[7]  Sato R,Takeuchi K,Ogiwara K, et al. Cloning heterologous expression and localization of a novel crystal protein gene from Bacillus thuringiensis serovar japonensis strain Buibui toxic to scarabaeid insects[M]. Curr Microbiol, 1994, 28, 15-19.
[8]  Asano S,Yamashita C, Iizuka T, et al. A strain of Bacillus thuringiensis subsp galleriae containing a novel cry8 gene highly toxic to Anomala cuprea ( Coleoptera: Scarabaeidae)[J]. Biol Control, 2003, 28( 2) : 191-196.
[9]  Schnepf E,Crickmore N,Van Rie J,et al. Bacillus thuringiensis and its pesticidal crystal proteins[J]. Microbiol Mol Biol Rev, 1998, 62: 775-806.
[10]  冯书亮,任国栋. 苏云金芽胞杆菌HBF-1 及其在有害金龟治理中的应用[M]. 北京: 中国农业科学技术出版社, 2005.
[11]  Yu H,Zhang J,Huang D F, et al. Characterization of Bacillus thuringiensisstrain Bt185 toxic to the asian cockchafer: Holotrichia parallela[J]. Current Microbiology, 2006, 53: 13-17.
[12]  Shu C,Liu R,Wang R, et al. Improving toxicity of Bacillus thuringiensis strain contains the cry8Ca gene specific to Anomala corpulenta larvae[J]. Current Microbiology, 2007, 55: 492-496.
[13]  Shu Changlong,Yan Guixin,Wang Rongyan, et al. Characterization of a novel cry8 gene specific to Melolonthidae pests: Holotrichia oblita and Holotrichia parallela[J]. Appl Microbiol Biotechnol, 2009, 84: 701-707.
[14]  刘兴龙,程林友,李天龙,等. Bt 杀虫蛋白基因的克隆、表达和活性[J]. 华北农学报, 2008, 23( 2) : 1-4.
[15]  Sambrook J,Fritschand E F,Maniatis T. Molecular cloning, A laboratory manual [M]. 2nd edition. New York: Cold Spring Harbor Laboratory Press, 2002.
[16]  任莹博,宋福平,陈中义,等. 苏云金芽孢杆菌cry1Aa14 基因的分离、克隆及其表达[J]. 农业生物技术学报, 2004, 12 ( 2) : 188-191.
[17]  Narva K E,Payne J M,Schwab G E, et al. European Patent Office no. EP 0462721[P]. 1991.
[18]  中国科学院微生物所. 伯杰氏细菌鉴定手册[M]. 北京: 科学出版社, 1984.
[19]  Nickerson K W, Julian St G,Bulla L A. Physiology of sporeforming bacteria associated with insects: radiorespirometric survey of carbohydrate metabolism in the 12 serotypes of Bacillus thuringiensis[J]. Appl Microbiol, 1974, 28: 129-132.
[20]  ZHAO J Z,CAO J,LI Y X, et al. Transgenic Plants Expressing Two Bacillus thuringiensis Toxins Delay Insect Resistance Evolution [J]. Nature Biotechnology,2003, 21( 12) : 1493-1497.

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133