表达绿色荧光蛋白的MVA重组毒株的构建及筛选

doi:10.16431/j.cnki.1671-7236.2019.07.024

Abstract

Abstract:

To construct and screen the recombinant modified vaccinia virus Ankara (MVA) strain expressing green fluorescent protein (GFP),primers were designed and synthesized.GFP gene was amplified by PCR and inserted into the ORF086-087 site (70303-70304 bp) of MVA based on the principle of homologous recombination.Fluorescence microscope was used to select single plaque with GFP as the reporter gene,and the recombinant strain was identified by fluorescence technique,PCR and Western blotting.The results showed that a large number of single plaques expressing GFP could be observed under fluorescence microscope,GFP gene successfully intergrated into recombinant strain MVA-GFP genome.Western blotting result showed that GFP was successfully expressed in infected cell.In this study,the recombinant strain MVA-GFP expressing GFP was constructed by genetic engineering technology,which provided materials for further screening unmarked recombinant strain and vaccines by inserting foreign target genes into GFP site.

Key words: green fluorescent protein (GFP); vaccinia virus MVA strain; recombinant strain

CLC Number:

S855.3

HUANG Caiyun, ZHAO Zhixun, ZHU Xueliang, WANG Zhanhong, WU Xiangcao, WU Guohua, ZHANG Zhidong, ZHANG Qiang. Construction and Screening of GFP Recombinant MVA Strain[J]. , 2019, 46(7): 2079-2087.

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Construction and Screening of GFP Recombinant MVA Strain

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