鼠疫耶尔森菌csf1基因的克隆表达及其产物免疫学评价

王鹏; 石国祥; 王海滨; 俞东征; 张建中

Abstract

王鹏,石国祥,王海滨,俞东征,张建中.鼠疫耶尔森菌csf1基因的克隆表达及其产物免疫学评价[J].Chinese journal of Epidemiology,2010,31(1):69-72

鼠疫耶尔森菌csf1基因的克隆表达及其产物免疫学评价

Clone expression of the csf1 gene of Yersinia pestis and immunological evaluation on recombinant FI antigen

Received:July 30, 2009

DOI：

KeyWord: 鼠疫耶尔森菌重组Fl抗原 csf1基因克隆表达免疫学活性

English Key Word: Yersinia pestis Recombinant FI antigen csf1 gene Clone expression Immunological activity

FundProject:国家高技术研究发展计划(863计划)(2006AA224A7)

Author Name	Affiliation	E-mail
WANG Peng	Department of Diagnosis. National Institute for Communicable Disease Controf and Prevention, Chinese Centerfor Disease Control and Prevention, Beijing 102206, China
SHI Guoxiang	Zhejiang Provincial Center for Disease Control and Prevention
WANG Haibin	Department of Diagnosis. National Institute for Communicable Disease Controf and Prevention, Chinese Centerfor Disease Control and Prevention, Beijing 102206, China
YU Dongzheng	Department of Diagnosis. National Institute for Communicable Disease Controf and Prevention, Chinese Centerfor Disease Control and Prevention, Beijing 102206, China
ZHANG Jianzhong	Department of Diagnosis. National Institute for Communicable Disease Controf and Prevention, Chinese Centerfor Disease Control and Prevention, Beijing 102206, China	zhangjianzhong@icdc.cn

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Abstract:

目的表达具有免疫学活性重组F1抗原(rFl)，并以其构建检测鼠疫抗体胶体金试纸条。方法将去掉信号肽编码序列的c口朋基因片段与载体质粒pET32a(+)通过BamHI和Not I双酶切位点进行连接，将重组质粒[csf1-pET32a(+)]转化入BL21(DE3)中进行诱导表达，表达产物经亲和层析纯化，以纯化rFl及天然F1抗原制备双检测鼠疫抗体胶体金标试纸条，并对浙江省528份人血清标本进行检测。结果含csf1-pET32a(+)的BL21(DE3)，经诱导产生相对分子质量(M_r)约为35.5X 103的rFl融合蛋白；rFl融合蛋白检测鼠疫抗体的敏感性等同甚至超越天然F1抗原；在528份人血清标本的检测中，rFl与天然F1的符合率为97.9％(K=0.466)，有较好一致性。结论制备的rFl，具有良好免疫学活性，该rFl可替代天然F1抗原，用于鼠疫免疫学检测。

English Abstract:

0bjective To get recombinant FI antigen(rFl)and to construct the detection dipstick of plague antibody.Methods The csf1 gene removing the signal peptide coding sequence was cloned into plasmid pET32a(+)by double.digested sites of BamHI and Not I.Recombinant plasmid csf1 -pET32a(+)was transformed into BL2l(DE3)and the rFl was expressed.Expression products were purified by affinity chromatography.Dual detection dipstick of plague antibody was constructed with purified rFl and natural F1.and evaluated with 528 human serum samples of Zhejiang province.Results The fusion protein rF l of 35.5 KD was expressed by BL21 slrains containing csf1 —pET32a(+).The sensitivity of rFl showed equivalent to or higller than the natural F1 antigen in detecting plague antibody.It seemed that there was a better consistency of 97.9％(K=0.466)when 528 human Sel'a was detected by rFl and natural F1.Conclusion We successfully extracted the rFl with good immunological activity that might be used to detecting Yersinia pestis.

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