[Cloning of bovine c-myc gene and its expression in skin fibroblast cells].
In order to construct a eukaryotic expression vector of bovine c-myc gene, the coding sequence (CDS) of c-myc gene was amplified from bovine primordial genital ridges by RT-PCR. The CDS was subcloned into pMD19-T vector, and then inserted into vector pIRES2-AcGFP1-Nuc. After confirmed by restriction enzyme digestion and sequencing, the recombined plasmid was transfected into skin fibroblast cells. RT-PCR and Western Blotting were used to detect the expression of c-myc mRNA and protein, respectively. The results show that the complete CDS of c-myc gene was cloned from fetal bovine primordial genital ridges. The eukaryotic expression vector of bovine c-myc gene was constructed and efficiently expressed in the skin fibroblast cells. The present study will lay a good foundation for further study of c-myc gene function and bovine induced pluripotent stem cells from somatic cells by defined factors.
College of Veterinary Medicine, Northwest Agriculture and Forestry University, Yangling 712100, China.
Jiajia
J
Xiao
Fengfeng
F
Zhang
Xianrong
X
Xiong
Yong
Y
Zhang
D000818:Animals; D002417:Cattle; D003001:Cloning, Molecular; D005333:Fetus / Q000166:cytology; D005347:Fibroblasts / Q000166:cytology*; D005822:Genetic Vectors / Q000235:genetics*; D016271:Proto-Oncogene Proteins c-myc / Q000096:biosynthesis* / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D012867:Skin / Q000166:cytology*; D014162:Transfection
D016271:Proto-Oncogene Proteins c-myc; D012333:RNA, Messenger; D011994:Recombinant Proteins