CAJ | 학술논문

为研究猪葡萄球菌表皮脱毒素 Exh C 基因的分子特征及其原核表达产物的免疫学活性，利用PCR 技术从猪葡萄球菌广东增城分离株（GDZC 株）扩增出表皮脱落毒素 Exh C 基因，测序与序列比较分析显示，其编码区全长为837 bp，编码279个氨基酸，与国内外猪葡萄球菌和松鼠葡萄球菌表皮脱落毒素 Exh C 基因的核苷酸同源性、氨基酸同源性均为100％。将 Exh C 基因克隆于 pET-28a （＋）表达载体中，经测序正确后，重组质粒转入大肠埃希菌 BL21（DE3）进行诱导表达，表达产物经 SDS-PAGE 鉴定和 Western blot分析。表达的融合蛋白主要以包涵体形式存在，其分子质量大小约为32 ku，与预期大小一致。Western blot 分析表明，Exh C 重组蛋白能与猪葡萄球菌 GDZC 株的阳性血清发生特异性反应，说明表达的 GDZC株 Exh C 重组蛋白具有良好的免疫学活性。
위연구저포도구균표피탈독소 Exh C 기인적분자특정급기원핵표체산물적면역학활성，이용PCR 기술종저포도구균엄동증성분리주（GDZC 주）확증출표피탈락독소 Exh C 기인，측서여서렬비교분석현시，기편마구전장위837 bp，편마279개안기산，여국내외저포도구균화송서포도구균표피탈락독소 Exh C 기인적핵감산동원성、안기산동원성균위100％。장 Exh C 기인극륭우 pET-28a （＋）표체재체중，경측서정학후，중조질립전입대장애희균 BL21（DE3）진행유도표체，표체산물경 SDS-PAGE 감정화 Western blot분석。표체적융합단백주요이포함체형식존재，기분자질량대소약위32 ku，여예기대소일치。Western blot 분석표명，Exh C 중조단백능여저포도구균 GDZC 주적양성혈청발생특이성반응，설명표체적 GDZC주 Exh C 중조단백구유량호적면역학활성。
This research was intended to study the molecular characteristics and reactogenicity of prokary-otic expression products of exfoliative toxin Exh C gene in S.hyicus.The Exh C gene was amplified in GDZC strain of S.hyicus which came from Zengcheng of Guangdong province by PCR.Sequence analysis indicated that the coding region has 837 bp,to code 279 amino acid.The nucleotide homology and amino acid homology were 100% both with Exh C gene of S.hyicus and S.sciuri which came from abroad.The recombinant expression plasmid pET-28a (+)-Exh C was shifted to E.coli BL21(DE3)for inducible ex-pression.After the SDS-PAGE,fusion protein was as inclusion body,and its molecular mass was 32 ku as the expected.Western blot indicated that recombinant protein of Exh C can response specifically with posi-tive serum of GDZC strain,and indicated that it has good immunologic activity.