中华临床感染病杂志
中華臨床感染病雜誌
중화림상감염병잡지
CHINESE JOURNAL OF CLINICAL INFECTIOUS DISEASES
2012年
2期
81-84
,共4页
朱坚胜%阮仙利%汪志军%周林福%王昕昕%王丽珍%陈华忠%王赓歌
硃堅勝%阮仙利%汪誌軍%週林福%王昕昕%王麗珍%陳華忠%王賡歌
주견성%원선리%왕지군%주림복%왕흔흔%왕려진%진화충%왕갱가
重组蛋白质类%酶联免疫吸附测定%埃可病毒
重組蛋白質類%酶聯免疫吸附測定%埃可病毒
중조단백질류%매련면역흡부측정%애가병독
Recombinant proteins%Enzyme-linked immunosorbent assay%Echovirus
目的 表达埃可病毒30型VP1重组蛋白,建立埃可病毒特异性抗体的检测方法.方法 采用反转录聚合酶链反应(RT-PCR)方法获得埃可病毒VP1cDNA,克隆至原核表达载体pET44a中表达,并用纯化的重组蛋白检测血清中埃可病毒抗体.结果 表达的重组蛋白VP1相对分子质量约为95 000,经Western blot和酶联免疫吸附试验证实其具有良好的抗原性.结论 基因工程重组融合蛋白VP1可以作为诊断用抗原.
目的 錶達埃可病毒30型VP1重組蛋白,建立埃可病毒特異性抗體的檢測方法.方法 採用反轉錄聚閤酶鏈反應(RT-PCR)方法穫得埃可病毒VP1cDNA,剋隆至原覈錶達載體pET44a中錶達,併用純化的重組蛋白檢測血清中埃可病毒抗體.結果 錶達的重組蛋白VP1相對分子質量約為95 000,經Western blot和酶聯免疫吸附試驗證實其具有良好的抗原性.結論 基因工程重組融閤蛋白VP1可以作為診斷用抗原.
목적 표체애가병독30형VP1중조단백,건립애가병독특이성항체적검측방법.방법 채용반전록취합매련반응(RT-PCR)방법획득애가병독VP1cDNA,극륭지원핵표체재체pET44a중표체,병용순화적중조단백검측혈청중애가병독항체.결과 표체적중조단백VP1상대분자질량약위95 000,경Western blot화매련면역흡부시험증실기구유량호적항원성.결론 기인공정중조융합단백VP1가이작위진단용항원.
Objective To express VP1recombinant protein of Echovirus 30 (ECHO30) in E.coli BL21(DE3) and to develop an enzyme-linked immunoassay (EIA) based on the recombinant antigen for detecting antibodies to ECHO30.Methods The target VP1gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR).The PCR products of the gene were inserted into pET44a vector,and then expressed in E.coli BL21( DE3 ).The purified recombinant protein was used for the development of EIA.Results The molecular weight of the recombinant protein was 95 000,and the antigenicity of which was identified by Western blot and EIA.Conclusion The recombinant protein VP1has been successfully expressed and purified,which can be used as diagnostic antigen.
