Ovine Follistatin gene expression and functional analysis of follistatin domains.
- Author:
Ning ZHANG
1
;
Xuemei ZHANG
;
Mingjun LIU
;
Lixin TAN
Author Information
1. Key Laboratory of Animal Biotechnology of Xinjiang, Urumqi 830000, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Cells, Cultured;
Escherichia coli;
genetics;
metabolism;
Female;
Follistatin;
biosynthesis;
genetics;
Genetic Vectors;
genetics;
Lentivirus;
genetics;
metabolism;
Muscle, Skeletal;
cytology;
Ovary;
metabolism;
Prokaryotic Cells;
metabolism;
Protein Structure, Tertiary;
Recombinant Proteins;
biosynthesis;
genetics;
Sheep
- From:
Chinese Journal of Biotechnology
2010;26(8):1050-1056
- CountryChina
- Language:Chinese
-
Abstract:
In order to study ovine follistatin function, we amplified the total of 1038 base pair of ovine complete follistatin cDNA and cloned into pGEM-T vector by RT-PCR from ovine ovary RNA. After removal of the signal peptide it was subcloned into the pET41a to construct the prokaryotic expression vector, named pFSsig-. SDS-PAGE and Western blotting identified the 66 kDa product of the expression of follistatin cDNA. Based on the complete CDS sequence, we cloned follistatin N-terminal domain and domain 1 with PCR and inserted into pLEX-MCS lentiviral vector, named pFS-N+D1. After package and passage of lentivirus in 293T cells, and then infected sheep primary muscle cells (SPMC). The expression of FS N+D1 in SPMC was assayed by Western blotting. The cell growth curve of the infected SPMC and noninfected control cells displayed that FS N+D1 stablly transfected SPMC proliferated significantly faster than the control cells (P < 0.01). Our data inferred that ovine FS N+D1 domain had the function to stimulate sheep muscle cell growth.
