摘要
应用逆转录－聚合醇链反应（ＲＴ－ＰＣＲ）和基因重组技术，将从中国河北省慢性丙型肝炎患者血中扩增的丙型肝炎病毒（ＨＣＶ）ＮＳ５基因片段，克隆到表达载体ｐＫＰＬ－３ａ中，在大肠杆菌中以天然蛋白形式高效表达了ＮＳ５蛋白，表达量约占菌体总蛋白的２０．３％。Ｗｅｓｔｅｒｎｂｌｏｔ试验和酶免疫试验（ＥＩＡ）表明，此蛋白可与丙型肝炎患者血清发生特异性反应，
AfragmentofhepatitisCvirusNS5gene,amplifiedbyreversetranscriptasepolymerasechainreaction(RT-PCR)fromthebloodsampleofachronichepatitisCpatientinHebeiprovince,China.wasclonedintoanexpcessionvectorpKPL-3a.NS5proteinwasexpressedheavilyinE.coliinanon-fusedformanditaccountedfor20.3%* oftotalbacterialpro-tein.WesternblotandenzynneimmunoassayanalysisshowedthatthisexpressedproteincouldbespecificallyrecognizedbytheserafrompatientswithhepatitisC.Thenon-fusedNS5pmteinexpressedinthislaboratorymayprovidesastartingpointforthestudyonthemechanismofHCVreplication,forthepreparationofmonoclonalandpolyclonalantilodiestoNS5protein,andforsettingupthethirdgenecationAinti-HCVdiagnostic.kitinourcountry.
出处
《中华实验和临床病毒学杂志》
CAS
CSCD
1996年第3期254-257,共4页
Chinese Journal of Experimental and Clinical Virology
