Construction and identification of soluble cluster of differentiation antigen 40 ligand prokaryotic expression vector
ZHANG Zhiqiang
QIAO Luxin
WU Tao
LIU Dongjie
JI Le
CHEN Dexi
WU Gang
Abstract：Objective To construct containing human soluble cluster of differentiation anti-gen 40 ligand ( sCD40L) recombinant plasmid pGEX-4T-sCD40L. Methods The gene fragment of human leukocyte sCD40L was amplified by reverse transcription-polymerase chain reaction ( RT-PCR) . The fragment was ligated into T vector and ligated with 5-bromo-4-chloro-3-indolyl-β-D-ga-lactoside ( X-gal ) and isopropyl-β-D-thiogalactopyranoside ( IPTG ) . The positive clones were screened by IPTG. The positive clones were digested with BamHⅠand XhoⅠ, then ligated with pGEX-4T prokaryotic expression vector, and identified by restriction enzyme digestion and sequen-cing. Results The target DNA fragment of 474 bp was amplified by the complementary DNA ( cDNA) of L02 cells. The recombinant plasmid pGEX-4T-sCD40L was constructed by ligating the target frag-ment with T vector and screening for blue-white. The recombinants were obtained by transformation and screening, and the sequences were confirmed by restriction enzyme digestion and gene sequen-cing. Conclusion The prokaryotic expression vector pGEX-4T-sCD40L has been successfully con-structed, which lays a foundation for further study on the role of sCD40L in the diagnosis of hepato-cellular carcinoma ( HCC) .
Keywords：Soluble cluster of differentiation antigen 40 ligand ( sCD40L)Prokaryotic ex-pression vectorFusion protein
Publication Date：2017-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：4( 12-15 )