Packaging of the recombinant lentivirus of iRhom2 and its mutant and establishment of a stable iRhom2-expressing cell line
10.3969/j.issn.1005-4847.2015.06.010
- VernacularTitle:iRhom2及其突变基因重组慢病毒的包装和稳定表达细胞系的建立
- Author:
Ying YOU
;
Yunan MA
;
Lin ZENG
;
Zhaozeng SUN
- Publication Type:Journal Article
- Keywords:
iRhom2;
Lentivirus vetor;
Packaging cell;
Vero cell line
- From:
Acta Laboratorium Animalis Scientia Sinica
2015;(6):597-601
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a stable transfection cell line of iRhom2 and its mutant through recombinant lentivirus infection.Methods The full-length gene of iRhom2 and its mutant were cloned into the lentivirus vector Lenti-OE-Flag, and got recombinant lentiviral vector of Lenti-OE-iRhom2 and Lenti-OE-iRhom2mut.The constructed recombi-nant lentivirus vectors were transfected into HEK-293T packaging cells to obtain the recombinant virus.Vero cells were in-fected with recombinant virus.The stable expressing cell lines were obtained by pressure screening with puromycin. Results The recombinant lentivirus vectors were constructed and the recombinant virus was obtained.The stable express-ing cell lines were obtained using virus infection and the protein expression was testified with Western blotting.Conclu-sions Stable iRhom2-expressing Vero cell line and its mutant are achieved by recombinant lentivirus infection.It paves the way for future study on biological functions and mechanism of iRhom2.
