中国现代医生
中國現代醫生
중국현대의생
CHINA MODERN DOCTOR
2014年
14期
69-72,75
,共5页
王志宏%丁莹莹%冯娇娇%李连青%潘卫%郭存九
王誌宏%丁瑩瑩%馮嬌嬌%李連青%潘衛%郭存九
왕지굉%정형형%풍교교%리련청%반위%곽존구
柯萨奇病毒A组16型%VP1蛋白%原核表达%酶联免疫吸附
柯薩奇病毒A組16型%VP1蛋白%原覈錶達%酶聯免疫吸附
가살기병독A조16형%VP1단백%원핵표체%매련면역흡부
Coxsackievirus group A type 16(CA16)%VP1 protein%Peokaryotic expression%Enzyme-linked immunosor-bent assay
目的:构建重组质粒柯萨奇病毒A组16型(coxsackievirus group A type 16,CA16)VP1,并进行原核表达及鉴定。方法用PCR方法扩增CA16 VP1序列,构建其重组表达质粒pET21b-CA16 VP1,并转入大肠杆菌E. coli BL21(DE3)进行诱导表达及纯化,SDS-PAGE鉴定表达及纯化产物,间接ELISA方法检测重组蛋白CA16 VP1的抗原性和有效性。结果成功原核表达并纯化了CA16 VP1蛋白,可与小鼠抗CA16病毒血清特异性结合,与太原市老年人群中部分血清存在高反应性。结论 CA16 VP1蛋白获得成功表达,ELISA检测具有良好的抗原性和有效性,为CA16诊断试剂盒的研究奠定基础。
目的:構建重組質粒柯薩奇病毒A組16型(coxsackievirus group A type 16,CA16)VP1,併進行原覈錶達及鑒定。方法用PCR方法擴增CA16 VP1序列,構建其重組錶達質粒pET21b-CA16 VP1,併轉入大腸桿菌E. coli BL21(DE3)進行誘導錶達及純化,SDS-PAGE鑒定錶達及純化產物,間接ELISA方法檢測重組蛋白CA16 VP1的抗原性和有效性。結果成功原覈錶達併純化瞭CA16 VP1蛋白,可與小鼠抗CA16病毒血清特異性結閤,與太原市老年人群中部分血清存在高反應性。結論 CA16 VP1蛋白穫得成功錶達,ELISA檢測具有良好的抗原性和有效性,為CA16診斷試劑盒的研究奠定基礎。
목적:구건중조질립가살기병독A조16형(coxsackievirus group A type 16,CA16)VP1,병진행원핵표체급감정。방법용PCR방법확증CA16 VP1서렬,구건기중조표체질립pET21b-CA16 VP1,병전입대장간균E. coli BL21(DE3)진행유도표체급순화,SDS-PAGE감정표체급순화산물,간접ELISA방법검측중조단백CA16 VP1적항원성화유효성。결과성공원핵표체병순화료CA16 VP1단백,가여소서항CA16병독혈청특이성결합,여태원시노년인군중부분혈청존재고반응성。결론 CA16 VP1단백획득성공표체,ELISA검측구유량호적항원성화유효성,위CA16진단시제합적연구전정기출。
Objective To construct the recombinant plasmid of coxsackievirus group A type 16 (CA16)VP1,express in E.coli and identify expressed product. Methods The CA16 VP1 gene was amplified by PCR,The constructed recombi-nant plasmid pET21b-CA16 VP1 was transformed into E.coli BL21(DE3) for expession and purification,expressed and purified product was identified by SDS-PAGE,Indirect ELISA tested Antigenicity and validity of recombinant proteins CA16 VP1. Results The coxsackievirus group A type 16 VP1 protein was expressed in peokaryotic cells and purified, which showed specific reaction with mice antiserum against CA16 virus,high reactivity with part serums of the elderly population in Taiyuan city. Conclusion CA16 VP1 protein was successfully expressed, had good Antigenicity and va-lidity,which will lay a foundation for study on the CA16 diagnostic kits.