Purification of p24 protein expressed in Bacillus subtilis and evaluation of its immunogenicity in mice

Abstract

p24 protein is a component of the HIV particle capsid. It plays an essential role in HIV to infect into the host cell and in the cycle life of virus. Therefore, this protein can be used in the orientative study “to create and produce HIV’s vaccine”. This study created the new Bacillus subtilis strain which expressed p24 protein. B. subtilis a safety and non-toxic bacteria strain for humans and animals, has system expression to allow over expression recombinant protein up to 10-30 % of total proteins. Plasmid pHT1537 was cloned successfully, containing lysSN-6his-gagp24 gene to encode p24 protein fused with LysSN protein and to allow the expression of p24 protein in B. subtilis by IPTG inducer. The target protein in the cell was cheked by SDS-PAGE. The p24 fused protein was parified from His Trap column which contained Ni2+. Evaluation of the ability to produce antibody against p24 protein in mice by ELISA and Western blot was caried out.

N‐terminal LysSN‐His‐tag improves the production of intracellular recombinant protein in Bacillus subtilis

“…[30][31][32][33] Our latest With the objective of developing a double fusion tag that can both purify and increase the expression level of the fusion target protein, we have used LysSN-6xHis-tag in many previous studies for protein expression and purification in B. subtilis, such as casein kinase 1 (CK1 D), 34 heat-labile enterotoxin B subunit (LTB), 35 nonreceptor tyrosine kinase protein (cSrc), 36 celA, 21 Listeriolysin O (LLO), 37 protein of the HIV particle capsid (P24). 38 So far, there has been no specific evaluation of the effect of LysSN-6xHis on the expression level of the target protein compared with His-tag without LysSN. In this study, we evaluated the impact of LysSN-6xHis on the expression levels of two groups of genes: high-expression genes (gfp+ and bgaB) and lowexpression genes (egfp) in B. subtilis.…”