ShRNA against NSP4 gene inhibits the proliferation of bovine rotavirus in vitro.
- Author:
Fang-yuan CHEN
1
;
Hong-mei WANG
;
Shao-hua YANG
;
Jian-ming WU
;
Yun-dong GAO
;
Xiao LIU
;
Hong-jun YANG
;
Chang-fa WANG
;
Ji-feng ZHONG
;
Li-qun WANG
;
Hong-bin HE
Author Information
1. Life College of Northeast Agricultural University, Harbin 150030, China. chenfangyuan.1983@163.com
- Publication Type:Journal Article
- MeSH:
Animals;
Base Sequence;
Cattle;
Cell Line;
DNA, Recombinant;
genetics;
Glycoproteins;
deficiency;
genetics;
Molecular Sequence Data;
Plasmids;
genetics;
RNA, Small Interfering;
genetics;
Rotavirus;
genetics;
physiology;
Toxins, Biological;
genetics;
Viral Load;
genetics;
Viral Nonstructural Proteins;
deficiency;
genetics;
Virus Replication;
genetics
- From:
Chinese Journal of Virology
2010;26(3):244-248
- CountryChina
- Language:Chinese
-
Abstract:
Based on the NSP4 sequence of bovine rotavirus (BRV), the shRNA was designed and synthesized, and a shRNA recombinant lenti-virus vector RNAi-H1-89 was constructed. The recombinant RNAi-H1-89 Lenti-virus was packaged by transfecting the 293T cell with the recombinant vector RNAi-H1-89 and two helper plasmids using lipofectamine, and then used to infect MA104 cells. The MA104 cells were further infected with BRV strain G6 24h post-infection, with the LacZ shRNA recombinant lenti-virus as control. Thirty-six hours later, the CPE of the infected cells was observed under microscope, shRNA of NSP4 gene inhibited CPE in MA104 cell; the shRNA against NSP4 gene also inhibited NSP4 gene expression by RT-PCR, The virus titer in the cell culture supernatant was significant lower compared with the control group. The above results showed that RNAi-H1-89 against NSP4 gene could specifically silence NSP4 gene expression, and inhibit the proliferation of BRV.
