[1]章美娟,胡治强,夏瑀培,等.类鼻疽杆菌Ⅵ型分泌系统蛋白Hcp1的重组表达及免疫学性质鉴定[J].第三军医大学学报,2020,42(23):2296-2301.
 ZHANG Meijuan,HU Zhiqiang,XIA Yupei,et al.Recombinant expression and immunological characterization of hemolysin coregulated protein 1, Burkholderia pseudomallei type Ⅵ secretion system protein[J].J Third Mil Med Univ,2020,42(23):2296-2301.
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类鼻疽杆菌Ⅵ型分泌系统蛋白Hcp1的重组表达及免疫学性质鉴定(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
42卷
期数:
2020年第23期
页码:
2296-2301
栏目:
基础医学
出版日期:
2020-12-15

文章信息/Info

Title:
Recombinant expression and immunological characterization of hemolysin coregulated protein 1, Burkholderia pseudomallei type Ⅵ secretion system protein
作者:
章美娟胡治强夏瑀培袁思琪闫晶敏饶承龙李倩杨文波毛旭虎
陆军军医大学(第三军医大学)药学与检验医学系临床微生物及免疫学教研室
Author(s):
ZHANG Meijuan HU Zhiqiang XIA Yupei YUAN Siqi YAN Jingmin RAO Chenglong LI Qian YANG Wenbo MAO Xuhu

Department of Clinical Microbiology and Immunology, College of Pharmacy and Medical Laboratory, Army Medical University (Third Military Medical University), Chongqing, 400038, China

关键词:
类鼻疽杆菌Ⅵ型分泌系统蛋白溶血素共调节蛋白1重组表达
Keywords:
Burkholderia pseudomallei type Ⅵ secretion system protein hemolysin coregulated protein 1 recombinant expression
分类号:
R378.99; R394-33; R446.62
文献标志码:
A
摘要:

]目的重组表达类鼻疽杆菌Ⅵ型分泌系统溶血素共调节蛋白1(hemolysin coregulated protein 1,Hcp1),并对其免疫学性质进行鉴定。方法利用DNA重组技术以 pET-28a表达系统在E.coli BL21(DE3)中重组表达Hcp1;通过亲和层析制备高纯度 Hcp1蛋白;采用腹腔注射免疫小鼠制备抗Hcp1血清,ELISA检测抗体滴度,免疫印迹及免疫荧光分析抗体的免疫反应性。结果成功获得纯度达95%的重组Hcp1蛋白,该蛋白免疫小鼠后获得的抗血清具有较好的特异性和较高的抗体滴度(1∶512 000),表现出良好的免疫原性和免疫反应性。结论成功制备了具有生物学活性的Hcp1重组蛋白及其抗血清。 

Abstract:

ObjectiveTo express recombinant hemolysin coregulated protein 1 (Hcp1) of type Ⅵ secretion system (T6SS) of Burkholderia pseudomallei and identify its immunological properties. MethodsThe recombinant Hcp1 protein was expressed by using DNA recombination technology through pET-28a expression system in E.coli BL21(DE3), and then purified by affinity chromatography. Then 6-week-old BALB/c mice were immunized with the obtained Hcp1 via intraperitoneal injection, the serum samples were collected to detect the titers of anti-Hcp1 specific antibodies by ELISA. Meanwhile, the immune response against antigen-specific antibodies was evaluated by Western blotting and immunofluorescence assay. ResultsThe recombinant Hcp1 protein was successfully obtained, with a purity of 95%. The obtained recombinant protein stimulated the antisera of good specificity and high titer (1∶512 000), with good immunogenicity and immunoreactivity. ConclusionThe recombinant protein Hcp1 and its antisera are successfully prepared. 

更新日期/Last Update: 2020-12-04