羊口疮病毒ORF059基因的真核表达研究

doi:10.16431/j.cnki.1671-7236.2016.09.011

Abstract

Abstract:

The study was aimed to clonie and express ORF059 gene of Orf virus (ORFV).The genome of ORFV HCE attenuated vaccine strain was extracted and used as the template.Based on the referenced gene sequences of ORF059 gene on GenBank,the specific primers were designed,the fragment of ORF059 gene was amplified by PCR.After being purified,ORF059 gene was inserted into pEGFP-N1 vector to construct recombinant plasmid pEGFP-ORF059-N1.After being sequenced,the recombinant plasmid pEGFP-ORF059-N1 was transfected into NIH3T3 cells with liposomes.The expression of fusion protein was identified by the fluorescence microscope observation and Western blotting.The results showed that the ORF059 gene was 1 005 bp.The recombinant plasmid pEGFP-ORF059-N1 was constructed correctly.Green fluorescence could be observed in NIH3T3 cells transfected by pEGFP-ORF059-N1 under the fluorescence microscope.The expressed fusion protein was about 64 ku.The results laid a foundation for the further research on the mechanism of ORF059.

Key words: ORFV; ORF059 gene; eukaryotic expression; NIH3T3 cell

CLC Number:

Q786

ZHU Shu, PANG Feng, LI Guo-hua, LI Ya-ying, PENG Dong-mei, YANG Xiao-jian, NIE Xin, CAO Rui-yong, WANG Feng-yang, DU Li. Study on Eukaryotic Expression of ORF059 Gene of Orf Virus[J]. , 2016, 43(9): 2291-2295.

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Study on Eukaryotic Expression of ORF059 Gene of Orf Virus

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