Orf virus ORF047 gene eukaryotic expression and cell localization
CHEN Guo-hua, JIA Huai-jie, HE Xiao-bing, WANG Chun-yan, JIN Qi-wang, JIN Zhi-zhong
State Key Laboratory on Veterinary Etiological Biology/Ministry of Agriculture Key Laboratory of Veterinary Public Health/Lanzhou Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Lanzhou 730046,China
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.
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2018, Vol. 34 
