pcDNA3-HBsAg-p30-ROP2真核表达载体的构建与鉴定
pcDNA3-HBsAg-p30-ROP2진핵표체재체적구건여감정
Construction and identification of pcDNA3-HBsAg-p30-ROP2 expression vec-tor
  • 万方数据
    中国血吸虫病防治杂志 중국혈흡충병방치잡지
    CHINESE JOURNAL OF SCHISTOSOMIASIS CONTROL
    2014年 1期 46-50 ,共5页

    魏庆宽%王英婷%闫运琴%肖婷%李瑾%徐超%刘功振%刘娟美%仲维霞%尹昆%付斌%闫歌%黄炳成

    위경관%왕영정%염운금%초정%리근%서초%류공진%류연미%중유하%윤곤%부빈%염가%황병성

    弓形虫%表面抗原1(p30)%棒状体分泌抗原2(ROP2)%乙肝表面抗原%基因重组 弓形蟲%錶麵抗原1(p30)%棒狀體分泌抗原2(ROP2)%乙肝錶麵抗原%基因重組
    궁형충%표면항원1(p30)%봉상체분비항원2(ROP2)%을간표면항원%기인중조
    Toxoplasma gondii%Surface antigen 1(p30)%Rhoptry protein 2(ROP2)%HBsAg%Gene recombination
    目的：构建pcDNA3-HBsAg-p30-ROP2多基因重组表达载体，并对其进行初步鉴定。方法根据重组体pcDNA3-p30-ROP2酶切位点和乙型肝炎表面抗原（HBsAg）基因序列等因素设计合成引物，扩增HBsAg目的基因片段，再应用酶切、连接等分子生物学技术将HBsAg目的基因克隆至pcDNA3-p30-ROP2表达载体中。应用聚合酶链反应（PCR）初筛，再采用酶切、测序等技术对构建的重组表达载体pcDNA3-HBsAg-p30-ROP2进行鉴定。结果 PCR扩增出HBsAg基因片段，构建了pcDNA3-HBsAg-p30-ROP2多基因真核表达载体。PCR与酶切结果显示，该基因片段大小均与理论值相符；测序结果显示该重组表达载体包含了p30-ROP2和HBsAg目的基因的完整序列。结论成功构建了多基因重组表达载体pcDNA3-HBsAg-p30-ROP2，为进一步研究多基因核酸疫苗奠定了基础。
    목적：구건pcDNA3-HBsAg-p30-ROP2다기인중조표체재체，병대기진행초보감정。방법근거중조체pcDNA3-p30-ROP2매절위점화을형간염표면항원（HBsAg）기인서렬등인소설계합성인물，확증HBsAg목적기인편단，재응용매절、련접등분자생물학기술장HBsAg목적기인극륭지pcDNA3-p30-ROP2표체재체중。응용취합매련반응（PCR）초사，재채용매절、측서등기술대구건적중조표체재체pcDNA3-HBsAg-p30-ROP2진행감정。결과 PCR확증출HBsAg기인편단，구건료pcDNA3-HBsAg-p30-ROP2다기인진핵표체재체。PCR여매절결과현시，해기인편단대소균여이론치상부；측서결과현시해중조표체재체포함료p30-ROP2화HBsAg목적기인적완정서렬。결론성공구건료다기인중조표체재체pcDNA3-HBsAg-p30-ROP2，위진일보연구다기인핵산역묘전정료기출。
    Objective To construct a multi-gene recombinant pcDNA3-HBsAg-p30-ROP2 expression vector and identify it preliminarily. Methods According to recombinant pcDNA3-p30-ROP2 restriction sites,HBV HBsAg gene sequences of primers were designed and synthesized to amplify target fragment,and then cloned into pcDNA3-HbsAg-p30-ROP2 expression vector. Af-ter sequencing,it was identified finally by restriction enzyme digestion and other molecular biology techniques. Results HBV HBsAg gene segment was amplified by PCR and the multi-gene recombinant pcDNA3-HBsAg-p30-ROP2 expression vector was constructed and identified to be correct as theoretical values. The PCR and restriction enzyme digestion results showed that HBsAg and p30-ROP2 gene in recombinant plasmid were confirmed by DNA sequencing. Conclusion The multi-gene recombinant pcD-NA3-HBsAg-p30-ROP2 expression vector is successfully constructed.
    원문보기 원문다운로드 사이트로이동
저자의 최근 논문