TI:Prokaryotic expression and purification of human immunodeficiency virus p24 antigen 　
AU:HOU Jun , MAOPanyong , HONG Shiwen , HU Yan , YANG Jianyang , SHEN Honghui , ZHU Lei Department of Virology , The
AD:No. 302 Hospital of The People’s Liberation Army , Beijing 100039 , China
【Abstract】　Objective 　To express and purify the HIV p24 gene in E1coli cells ,and identify p24 antigen activity1 Methods 　The full length gene fragment of HIV p24 was amplified by PCR and inserted into the pRSET vector in order to construct the pRSET2p24 recombined vector1 After transforming into E1coil , the purified p24 protein was prepared by metal2ligand affinity chromatography ( IMAC) 1 The accuracy of inserted gene and activity , specificity of HIV p24 proteins were detected by two enzymes digestion technology , SDS2PAGE , Western Blot (WB) and ELISA1 Results 　The length of the HIV p24 gene fragment was 690 bp after digesting the recombinant plasmid T2p24 and pRSET2p24 with BamH Ⅰand Hind Ⅲ1 The expressed proteins had a single expected band of about 24 ×103 in SDS2PAGE1 The specificity and activity of p24 protein were tested by WB and ELISA1 Conclusion 　The HIV p24 sequence from HIV21 gene plasmid has been expressed in E1coil1 This protein possessed good specificity and activity1
【Key words】　HIV; 　Protein p24 antigen ; 　Gene expression ; 　Polymerase chain reaction