组成型表达人呼吸道合胞病毒F<sub>212-489</sub>蛋白乳酸乳球菌的构建

冯金; 洪愉; 毛普加; 黄芬; 井申荣; 曾韦锟

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组成型表达人呼吸道合胞病毒F_212-489蛋白乳酸乳球菌的构建

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- 组成型表达人呼吸道合胞病毒F_212-489蛋白乳酸乳球菌的构建
- Constitutive expression of human respiratory syncytial virus protein F_212-489 in Lactococcus lactis
- 陆军军医大学学报 2014年36卷第13期页码：1354-1358
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  中图分类号： R37;R392.1;R394-33
- 纸质出版：2014
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冯金,洪愉,毛普加,黄芬,井申荣,曾韦锟.组成型表达人呼吸道合胞病毒F212-489蛋白乳酸乳球菌的构建[J].陆军军医大学学报(原第三军医大学学报),2014,36(13):1354-1358.

　Feng Jin,Hong Yu,Mao Pujia,Huang Fen,Jing Shenrong,Zeng Weikun.Constitutive expression of human respiratory syncytial virus protein F212-489 in Lactococcus lactis[J].J Amry Med Univ (J Third Mil Med Univ),2014,36(13):1354-1358.
冯金,洪愉,毛普加,黄芬,井申荣,曾韦锟.组成型表达人呼吸道合胞病毒F212-489蛋白乳酸乳球菌的构建[J].陆军军医大学学报(原第三军医大学学报),2014,36(13):1354-1358. DOI：

　Feng Jin,Hong Yu,Mao Pujia,Huang Fen,Jing Shenrong,Zeng Weikun.Constitutive expression of human respiratory syncytial virus protein F212-489 in Lactococcus lactis[J].J Amry Med Univ (J Third Mil Med Univ),2014,36(13):1354-1358. DOI：

摘要

目的构建能表达呼吸道合胞病毒截短F1蛋白（F

212-489

）的乳酸乳球菌，并检测表达蛋白的免疫反应性，以评估此重组菌作为口服活菌疫苗的可行性。方法 PCR扩增绿色荧光基因egfp以及截短f1基因（f

212-489

），经TA克隆测序后，将目的片段构建至pMG36e载体，电转化至乳酸乳球菌中鉴定并表达，通过荧光显微镜检测绿色荧光蛋白EGFP的表达，通过SDS-PAGE和Western blot检测F

212-489

蛋白的表达。结果荧光显微镜观察到重组菌pMG36e-egfp/MG1363有绿色荧光表达，Western blot检测到重组菌pMG36e-f

212-489

/MG1363中F

212-489

蛋白的表达。结论构建的乳酸乳球菌能够组成型表达F

212-489

蛋白，其蛋白具备免疫反应性。

Abstract

Objective To construct a Lactococcus lactis system which could express the truncated F1 protein (F

212-489

) of human respiratory syncytial virus and to identify the immunoreactivity of the obtained protein in order to evaluate the possibility of recombinant bacteria as an oral vaccine. Methods Firstly

egfp and f

212-489

gene were amplified by PCR

and then the product was inserted into the pMD18T vector for sequencing. Secondly

recombinant bacteria pMG36e-egfp/MG1363 and pMG36e-f

212-489

/MG1363 were constructed. Finally

fluorescence microscopy

SDS-PAGE and

Western blotting were used to investigate the expressed protein. Results Expression of enhanced green fluorescent protein (EGFP) was found in the recombinant bacteria pMG36e-egfp/MG1363. F

212-489

 was detected in the recombinant bacteria by Western blotting. Conclusion The protein F

212-489

is correctly expressed by the recombinant Lactococcus lactis with its immunoreactivity.

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组成型表达人呼吸道合胞病毒F212-489蛋白乳酸乳球菌的构建

Constitutive expression of human respiratory syncytial virus protein F212-489 in Lactococcus lactis

DOI：

摘要

Abstract

关键词

Keywords

references

组成型表达人呼吸道合胞病毒F_212-489蛋白乳酸乳球菌的构建

Constitutive expression of human respiratory syncytial virus protein F_212-489 in Lactococcus lactis