乙型肝炎病毒S基因与截短C基因融合的穿梭表达载体的构建

Construction Shuttle Expression Vector Carry S Gene Fused with Truncated Core Gene from Hepatitis B Virus

研究目的是构建HBVS基因和截短C基因融合的胞壁型和分泌型大肠杆菌和分枝杆菌(E.coli-BCG)穿梭质粒。采用PCR方法从结核分枝杆菌MTB毒株H37Rv的基因中扩增出相对分子质量为19000的抗原胞壁区及其上游调控元件基因,克隆入穿梭载体pOLYG中。以含HBV基因组的质粒pCP10序列为模板,PCR扩增获得S基因片段Sw和C基因编码氨基端的部分基因片段Ct,克隆入胞壁型穿梭表达载体pCW和分泌型穿梭表达载体pDE22。经酶切和序列测定证实,胞壁型和分泌型载体pCW-Sw-Ct和pDE22-Sw-Ct构建成功。为进一步研究含S基因和截短C基因融合的重组BCG活疫苗奠定了基础。

Construct cellular wall and secreting expression vectors carry S gene fused with truncated core gene from hepatitis B virus that can shuttle between E.coli and BCG also. The gene encode 19 kD antigen and upstream adjust sequences was obtained by PCR from H37Rv MTB strains and cloned into a vector named pOLYG which can shuttle between E.coli and BCG to construct cellular wall vector pCW. Then S gene or truncated core gene which encode some N terminus amino acids was got from pCP10(carry HBV genosome) through PCR and dubbed Sw or Ct respectively. Both Sw and Ct was cloned into shuttle vector pCW or the secreting type named pDE22 in turn. Both restrictive enzyme cleavage and sequencing proved that pCW-Sw-Ct or pDE22-Sw-Ct that can shuttle between E.coli and BCG was constructed successfully. The foundation of recombinant BCG vaccine express S gene fused with truncated core gene was established.