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...

烟夜蛾幼虫中肠类钙粘蛋白基因cDNA片段的克隆与序列分析

DOI: 10.3321/j.issn:1000-7091.2007.01.036, PP. 152-155

Keywords: 烟夜蛾,类钙粘蛋白,基因克隆,序列分析

Full-Text   Cite this paper   Add to My Lib

Abstract:

类钙粘蛋白位于昆虫中肠刷状缘膜囊泡上,该蛋白作为Bt毒素的受体可以与其结合导致昆虫死亡,因此,昆虫对Bt毒素的抗性与类钙粘蛋白有密切关系。本研究以烟夜蛾五龄幼虫中肠的总RNA为模板,根据已经克隆的其他昆虫的类钙粘蛋白基因序列设计引物,利用RT-PCR技术扩增出了烟夜蛾类钙粘蛋白基因的cDNA片段,将其连接到pMD18-T载体,转化大肠杆菌后筛选阳性克隆并进行序列测定,测序得到的1335bp的片断编码444个氨基酸残基。序列分析表明,该氨基酸序列与棉铃虫和烟芽夜蛾类钙粘蛋白的一致性分别为95%和86%。研究结果对进一步研究昆虫对Bt毒素产生抗性的机制以及发展分子技术快速检测并监测田间昆虫抗性基因的产生奠定了基础。

References

[1]  安世恒, 郭线茹, 罗梅浩, 等. 烟夜蛾幼虫中肠CrylAc毒索受体蛋白cDNA片断的克隆和序列测定[J]. 华北农学报, 2005, 20(2): 84-88.
[2]  McGaughey W H. Insect resistance to the biological insecticide Bacillus thuringiensis[J]. Science, 1985, 229 (4709): 193-195.
[3]  Gunning R V, Dang H T, Kemp F C, et al. New resistance mechanism in Helicoverpa armigera threatens transgenic crops expressing Bacillus thuringiensis CrylAc toxin[J]. Appl Environ Microbiol, 2005, 71(5): 2558-2563.
[4]  Gahan L J, Gould F, Heckel D G. Identification of a gene associated with Bt resistance Heliothis virescenss[J]. Science, 2001, 293(5531): 857-860.
[5]  Hua G, Jurat-Fuentes J L, Adang M J. Bt-R1a extracellular cadherin repeat 12 mediates Bacillus thuringiensis Cry1Ab binding and cytotoxicity[J]. J Biol Chem, 2004, 279 (27): 28051-28056.
[6]  Dorsch J A, Candas M, Griko N B, et al. Cry1A toxins of Bacillus thuringiensis bind specifically to a region adjacent to the membrane-proximal extracellular domain of BT-R (1) in Mandluca sexta: involvement of a cadherin in the entomopathogenicity of Bacillus thuringiensis[J]. Insect Biochem Mol Biol, 2002, 32 (9): 1025-1036.
[7]  Morin S, Biggs R W, Sisterson M S, et al. Three cadherin alleles associated with resistance to Bacillus thuringiensis in pink bollworm[J]. PNAS, 2003, 100(9): 5004-5009.
[8]  Francis B R, Bulla L A Jr. Further characterization of BTR1, the cadherin-like receptor for Cry1Ab toxin in tobacco hornworm (Manduca sexta) midguts[J]. Insect Biochem Mol Biol, 1997, 27(6): 541-550.
[9]  Xu X, Yu L, Wu Y. Disruption of a cadherin gene associated with resistance to Cry1Ac delta-endotoxin of Bacillus thuringiensis in Helicoverpa armigera[J]. Appl Environ Microbiol, 2005, 71 (2): 948-954.
[10]  Nagamatsu Y, Toda S, Koike T, et al. Cloning, sequencing and expression of the Bombyx mori receptor for Bacillus thuringiensis insecticidal CryIA (a) toxin[J]. Biosci Biotechnol Biochem, 1998, 62 (4): 727-734.
[11]  Vadlamudi R K, Weber E, Ji I, et al. Cloning and expression of a receptor for an insecticidal toxin of Bacillus thuringiensis[J]. J Biol Chem, 1995, 270 (10): 5490-5494.
[12]  王桂荣, 吴孔明, 梁革梅, 等. 棉铃虫中肠钙粘蛋白基因的克隆、表达及Cry1A结合区定位[J]. 中国科学C辑, 2004, 34(6): 537-546.
[13]  Xie R, Zhuang M, Ross L S, et al. Single amino acid mutations in the cadherin receptor from Heliothis virescenss affeet its toxin binding ability to Cry1 A toxins[J]. J Biol Chem, 2005, 280(9): 8416-8425.
[14]  Coates B S, Sumerford D V, Hellmich R L, et al. Sequence variation in the cadherin gene of Ostrinia nubilalis: a tool for field monitoring[J]. Insect Biochem Mol Biol, 2005, 35 (2): 129-139.

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133