Please wait a minute...

中国生物工程杂志

China Biotechnology
China Biotechnology  2009, Vol. 29 Issue (05): 1-5    DOI:
    
The interaction between Human Trim5α Chimerae and HIV-1gag Protein in Vitro
Download: HTML   PDF(475KB) HTML
Export: BibTeX | EndNote (RIS)      

Abstract  

Objective: To express and purify the TRIM5α chimaera[TRIM5α-H(R328- 332)] protein and to explore the interaction between the TRIM5α H(R328- 332)and HIV-1gag. Methods: The plasmid pET28aTRIM5α H(R328-332) was transformed to E.coli BL21 (DE3) strain , and the expression of TRIM5α H(R328-332) protein was induced by IPTG,purified with Ni2+ chromatography.The expression and purification of TRIM5α H(R328-332) were analyzed by SDS-PAGE and Western blot,and the interaction between TRIM5α-H(R328-332) and HIV-1gag was detected by co-immunoprecipitation, His pull-down and ELISA. Results:The recombinant plasmid pET28aTRIM5α H(R328-332) was successfully expressed in E.coli. The results showed that the purified full length TRIM5α-H(R328-332) interacted with HIV-1gag protein. Conclusion: The human TRIM5α-chimaera was expressed successfully in vitro, and the study demonstrates that the human TRIM5α-chimaera interacts with HIV-1 gag in vitro.



Received: 14 January 2009      Published: 06 May 2009
ZTFLH:  Q789  
Cite this article:

MENG Xiang-Beng- Li-Xiu-Yang- Sun-Han-Xiao- Mo-Xue-Mei. The interaction between Human Trim5α Chimerae and HIV-1gag Protein in Vitro. China Biotechnology, 2009, 29(05): 1-5.

URL:

https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/Y2009/V29/I05/1

[1] Hogan W,Schubert D,LaBonte J,et a1.Speciesspecific postentry barriers to primate immunodeficiency virus infection.J Viml,1999,73(12):1020~1028 [2] Owens C M,Yang P C,Gotflinger H,et al.Human and simian immunodeficiency virus capsid proteins are major viral determinants of early postentry replication blocks in simian cells.J Virol,2003,77(1):726~731 [3] Yap M W,Nisole S,Stoye J P.A single amino acid change in the SPRY domain of human TRIM5α leads to HIV-1 restriction.CurrBioZ,2005,15(1):73~78 [4] Khazaeli M B, Conry R M, LoBuglio A F. Human response to monoclonal antibodies. J Immunother,1994, 15: 42~52 [5] Matthew Stremlau, Michel Perron, Sohanya Welikala.Species-specific variation in the B30.2(SPRY)domain of TRIM5α determines the potency of human immunodeficiency virus restriction.Journal of Virology, 2005, 79( 5):3139~3145 [6] 饶海林,张珂,曾海鹏,等.大肠杆菌表达的人IL-6复性条件研究. 微生物学免疫学进展,2007,35(3):27~29 Rao H L,Zhang K,Zeng H P, et al. Prog in Microbiol Immunol,2007,35(3): 27~29 [7] 金腾川,张瑞军,魏海明,等.重组人白细胞介素-18的高浓度复性、纯化和活性检测.中国生化药物杂志,2004,25(2):67~71 Jin T C, Zhang R J, Wei H M,et al.Chinese Journal of Biochemical Pharmaceutics,2004,25(2):67~71 [8] Hassan Javanbakht, Wen Yuan, Darwin F. Characterization of TRIM5α TRIMerization and its contribution to human immunodeficiency virus capsid binding.Virology,2006, 353:234~246 [9] 李帷.生物超分子体系.北京:化学工业出版社,2003:225226 Li W. Biological Supramolecular System. Beijing:Chemical industry press, 2003:225~226 [10] 柴智,罗旭光,徐钧,等.乙型肝炎表面抗原单克隆抗体的制备及其应用.中国生物工程杂志,2008, 28(1): 70~75 Chai Z,Luo X G,Xu J,et al. China Biotechnology, 2008, 28(1): 70~75

No related articles found!