[Site-directed mutagenesis of MSTN gene by gene targeting in Qinchuan cattle].
Myostatin, a member of the transforming growth factor beta (TGF-beta) family, is a negative regulator for muscle growth. Loss of the function of this gene is associated with the phenotype described as "double muscling", an extreme form of muscle development characterized by a large increase in muscle mass. Two replacement vectors, pA2T-Mstn4.0 and pA2T-Mstn3.2, were constructed, linearized, and transfected into the bovine fetal fibroblasts through electroporation. 170 drug-resistant cell colonies were obtained in cell culture medium containing 600 microg/mL G418 and 50 nmol/L GCV. Targeted homologous integration occurred in colony No. 58 as identified by PCR, and the targeted colony was further confirmed by sequencing and Southern blotting. This suggested that one allele of myostatin was successfully mutagenized in bovine fetal fibroblasts.
Key Laboratory of Animal Reproductive Endocrinology & Embryo Engineering, Ministry of Agriculture, College of Veterinary Medicine, Northwest A & F University, Yangling 712100, China.
Yonggang
Y
Liu
Song
S
Hua
Jie
J
Lan
Yongli
Y
Song
Yulong
Y
He
Fusheng
F
Quan
Yong
Y
Zhang
D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002681:China; D018274:Electroporation; D005333:Fetus; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D018390:Gene Targeting; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed / Q000662:veterinary*; D055435:Myostatin / Q000235:genetics*
D055435:Myostatin