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...

铜绿假单胞菌含Pilz结构域蛋白基因的克隆及原核表达

, PP. 564-567

Keywords: 铜绿假单胞菌,c-di-GMP第二信使,Pilz结构域蛋白,基因克隆,原核表达载体构建

Full-Text   Cite this paper   Add to My Lib

Abstract:

目的克隆铜绿假单胞菌含Pilz结构域蛋白基因并构建其原核表达载体,为进一步研究这七种含Pilz结构域蛋白是否为第二信使c-di-GMP潜在的受体分子奠定实验基础。方法提取铜绿假单胞菌PAO1基因组DNA,用PCR方法从中扩增出七种功能未知的含Pilz结构域蛋白基因,将其克隆到原核表达载体pET32a中,在大肠杆菌BL21(DE3)中用IPTG诱导表达,表达产物通过十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)进行鉴定。结果成功扩增出七种含Pilz结构域蛋白基因并构建其原核表达载体,测序结果与NCBI数据库收录序列相一致。SDS-PAGE检验证明目的基因在大肠杆菌BL21中获得高效表达。结论从铜绿假单胞菌基因组中成功地克隆了七种含Pilz结构域蛋白基因,其构建的原核表达载体在大肠杆菌BL21中获得了高效表达。

References

[1]  CK Stover, XQ Pham, AL Erwin, et al. Complete genome sequence of Pseudomonas aeruginosa PAO1, an opportunistic pathogen [J]. Nature, 2000,406:959-964.
[2]  Kumar M, Chatterji D. Cyclic di-GMP: a second messenger required for long-term survival, but not for biofilm formation, in Mycobacterium smegmatis[J]. Microbiology, 2008, 154:2942-2955.
[3]  Ross P, Aloni Y, Weinbouse H, et al. Control of cellulose biosynthesis in Acetobacter xylinum: a unique quanyl oligonucleotide is the immediate activator of the cellulose synthase [J]. Carbobydrate Res,1986, 149: 101-117.
[4]  P Ross, H Weinhouse, Y Aloni, et al. Regulation of cellucose synthesis in Acetobacter xylinum by cyclic diguanylic acid [J]. Nature, 1987, 325: 279-281.
[5]  RomLing U, Amikam D. Cyclic di-GMP as a second messenger [J]. Curr Opin Microbiol, 2006,9:218-228.
[6]  Tamayo R, Pratt JT, Camilli A. Roles of cyclic diguanylate in the regulation of bacterial pathogenesis [J]. Annu Rev Microbiol, 2007, 61: 131-148.
[7]  Amikam D, Galperin MY. Pilz domain is part of the bacterial cdi-GMP binding protein [J]. Bioinformatics, 2006, 22:3-6.
[8]  Ryjenkov DA, Simm R, RomLing U, et al. The Pilz domain is a receptor for the second messenger c-di-GMP: the Pilz domain protein YcgR controls motility in enterobacteria [J]. J Biol Chem, 2006, 281: 30310-30314.
[9]  Merighi M, Lee VT, Hyodo M, et al. The second messenger bis-(30-50)-cyclic-GMP and its Pilz domain containing receptor Alg44 are required for alginate biosynthesis in Pseudomonas aeruginosa [J]. Mol Microbiol, 2007, 65:876-895.

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133