Orf virus ORF047 gene eukaryotic expression and cell localization
10.3969/j.issn.1002-2694.2018.00.015
- VernacularTitle:羊传染性脓疱病毒ORF047基因表达及其蛋白在细胞中的定位分析
- Author:
Guo-Hua CHEN
1
;
Huai-Jie JIA
;
Xiao-Bing HE
;
Chun-Yan WANG
;
Qi-Wang JIN
;
Zhi-Zhong JIN
Author Information
1. 中国农业科学院兰州兽医研究所/家畜疫病病原生物学国家重点实验室/农业部兽医公共卫生重点实验室
- Keywords:
contagious ecthyma;
ORF047 gene;
cell location
- From:
Chinese Journal of Zoonoses
2018;34(2):129-132
- CountryChina
- Language:Chinese
-
Abstract:
The object of study was to clone the gene of ORFV ORF047 and study the eukaryotic expression and cell localization,making the theoretical basis for the subsequest screening of protein that interact with ORF047.ORF047 gene was amplificated by the specifical primer from the DNA of ORFV using PCR,the length was 735 bp,compared with L1 published in NC-005336.1,the homologies of the nucleotide acid sequence and amino acid sequence were 98.8％ and 98.8％.In order to defined the expression and location of the ORF047 gene in cell,the recombinant plasmid pEGFP-ORF047 was constructed and transfected into 293T cell,after 36 h,the green fluorescence could be observed under fluorescence microscope,and 54 kD protein was detected by western bloting.The plasmid of pHcRed1-Nuc,pHcRed1-Mito and pHcRed1-ER with the recombinant plasmid of pEGFP-ORF047 was cotransfected to veroE6 cell respectively,that fusion protein of ORF047 was mainly located in the cytoplasm,a small amount in the mitochondriabyconfocal microscopy analysis.
