enJSRV囊膜蛋白及其受体的瞬时表达与绵羊绒毛膜滋养层细胞融合的诱导研究

doi:10.11843/j.issn.0366-6964.2015.11.003

Abstract

Abstract:

This paper aimed to study the possible role of endogenous jaagsiekte sheep retrovirus（enJSRV） envelope protein in sheep placenta formation process.Chorionic trophoblast cells from Mongolian sheep were cultured in vitro.RT-PCR technique was used to amplify enJSRV-env gene as well as the full-length gene of its receptor Hyal2.The genes were then directionally cloned into the eukaryotic expression vector pEGFP-C1.The electric transfection conditions of the chorionic trophoblast cells were optimized and the electrical transfer efficiency was measured.The constructed eukaryotic expression plasmids were transfected into chorionic trophoblast cells by electroporation，and transient expressions of enJSRV envelope protein and its receptor Hyal2 were observed under a fluorescence microscope.The activity of cell fusion in chorionic trophoblast cells under high expression and RNA interference silencing of enJSRV-env gene was also observed.The results showed that eukaryotic expression plasmids were successfully constructed and named as pEGFP-C1/enJSRV-env and pEGFP-C1/Hyal2，respectively.The best transfection conditions were pulse voltage 150 V，pulse time 5.0 ms，electric shock number for 2 times with 50 ms interval.The number of multinucleated cells were significantly increased in the trophoblast cells which co-transfected pEGFP-C1/Hyal2 with the transfection of pEGFP-C1/enJSRV-env plasmid and pEGFP-C1/enJSRV-env.We observed 2.3 and 1.8 multinucleated cells in average per field.The enJSRV-env had a certain role in promoting the cell fusion of chorionic trophoblast cells.This study provides an experimental basis for further exploring the structure and function of enJSRV envelope protein and the fusion mechanism of chorionic trophoblast cells.

CLC Number:

S826.2

ZHANG Yu-fei，SHI Jing，LIU Shu-ying. Study on the Transient Expression of enJSRV Envelope Protein and Its Receptor and the Induction of Trophoblast Cell Fusion in Sheep[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(11): 1924-1933.

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Study on the Transient Expression of enJSRV Envelope Protein and Its Receptor and the Induction of Trophoblast Cell Fusion in Sheep

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