[Envelope protein of Jaagsiekte sheep retrovious expressed in NIH3T3 cells promotes cell proliferation].
Objective To explore the influence of the exogenous Jaagsiekte sheep retrovious (exJSRV) envelope protein (Env) on NIH3T3 cell proliferation. Methods A recombinant plasmid pcDNA4/myc-His/exJSRV- env carrying exJSRV- env gene was constructed, and then the correctness of the recombinant plasmid was identified by PCR, restriction enzyme digestion and sequencing. The recombinant plasmid pcDNA4/myc-His/exJSRV- env was transiently transfected into NIH3T3 cells by Lipofectamine(TM) LTX. After the transfection of the recombinant plasmid, the expression of exJSRV- env was detected by reverse transcription PCR and Western blotting. The effect of Env on cell proliferation was investigated by CCK-8 assay and plate colony formation assay. Results The recombinant eukaryotic expression plasmid containing exJSRV- env was successfully constructed as identified by PCR, restriction enzyme identification and sequencing. After the recombinant plasmid was transiently transfected into NIH3T3 cells, reverse transcription PCR and Western blotting showed the expression of exJSRV- env , and Env promoted NIH3T3 cell proliferation significantly. Conclusion JSRV Env was expressed successfully in the NIH3T3 cells and promoted the proliferation of NIH3T3 cells.
College of Veterinary Medicine, Inner Mongolia Agricultural University, Huhhot 010018, China.
Fangyuan
F
DU
Inner Mongolia Sainuo Sheep Pasture Corporation, Hohhot 010040, China.
Dayong
D
Chen
College of Veterinary Medicine, Inner Mongolia Agricultural University, Huhhot 010018, China.
Yufei
Y
Zhang
College of Veterinary Medicine, Inner Mongolia Agricultural University, Huhhot 010018, China.
Xiaolin
X
Sun
College of Veterinary Medicine, Inner Mongolia Agricultural University, Huhhot 010018, China.
Wenqing
W
Guo
College of Veterinary Medicine, Inner Mongolia Agricultural University, Huhhot 010018; 3Ministry-of-Agriculture Key Laboratory of Clinical Diagnosis and Treatment Technology in Animal Disease, Huhhot 010018, China. *Corresponding author, E-mail: liushuying_imau@126.com.
Shuying
S
Liu
D000818:Animals; D030121:Betaretrovirus / Q000235:genetics* / Q000378:metabolism; D049109:Cell Proliferation; D015870:Gene Expression; D051379:Mice; D041681:NIH 3T3 Cells; D011648:Pulmonary Adenomatosis, Ovine / Q000503:physiopathology* / Q000821:virology*; D012756:Sheep; D014162:Transfection; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000378:metabolism
D014759:Viral Envelope Proteins