Contact Us Add to Favorite
 
NianQi Search Adv Search
33
研究论文 Current Issue | Archive | Adv Search |
Cloning, Expression and Purification of Chicken MHC I α-BirA Substrate Peptide (BSP) Sequence in Escherichia coli
LIU Guang-liang;TONG Tie-gang;WANG Qun;XIAO Yi-hong;MENG Qing-wen;WU Dong-lai
Harbin Veterinary Research Institute, Chinese  Academy of Agricultural  Sciences, Harbin 150001, China; 2. Graduate School,Chinese  Academy of Agricultural Sciences, Beijing 100081, China
Download: PDF (610 KB)   HTML (1 KB) 
Export: BibTeX | EndNote (RIS)      
Abstract  The gene of chicken MHC I α (BF2 ) was amplified from total RNA of peripheral blood by reverse transcription-polymerase chain reaction (RT-PCR) and cloned into pMD18-T. And then the PCR product which deleted the signal sequence at the 5' end in the BF2 gene and included BirA substrate peptide(BSP ) sequence at the 3' end was amplified from pMD18-T including BF2 gene by PCR amplification and subcloned into an expression vector pET-28a(+) named pET-BF2-BSP. The recombinant vector pET-BF2-BSP successfully expressed in the Escherichia coli strain BL21(DE3). The results of SDS-PAGE and Westernblot showed that the target protein was about 40 kD containing 6×His tag.  The overexpressed recombinant protein could be gained when the bacteria was at OD600nm of 0.8 to 1.2 induced for 2 to 4 h and 1.1 mmol/L IPTG. Additionally, the recombinant target protein was purified with a Ni2+NTA column including His-Bind Resin and high quality of recombinant protein  was recovered from the inclusion body. All these results indicated that high quality of recombinant protein of BF2-BSP was obtained, which will be contributed to the construction of chicken MHC peptide tetramer.
Key wordschicken MHC      cloning      BirA substrate peptide sequence      expression      purification     
Received: 13 December 2005     
ZTFLH:  S188  
Corresponding Authors: , WU Dong-lai   
Service
E-mail this article
Add to my bookshelf
Add to citation manager
E-mail Alert
RSS
Articles by authors
LIU Guang-liang
TONG Tie-gang
WANG Qun
XIAO Yi-hong
MENG Qing-wen
WU Dong-lai
Cite this article:   
LIU Guang-liang,TONG Tie-gang,WANG Qun, et al. Cloning, Expression and Purification of Chicken MHC I α-BirA Substrate Peptide (BSP) Sequence in Escherichia coli[J]. , 2006, 14(5): 662-668.
URL:  
http://journal05.magtech.org.cn/Jwk_ny/EN/      OR     http://journal05.magtech.org.cn/Jwk_ny/EN/Y2006/V14/I5/662
Copyright © Editorial Board of 农业生物技术学报
Supported by:Beijing Magtech