Myostatin gene targeting in cultured China Han ovine myoblast cells
L. Zhang,
X. Yang,
X. An,
Y. Chen
Affiliations
L. Zhang
State Key Laboratory for Agrobiotechnology, College of Biology, China Agricultural University, Yuanming Yuan West Road #2, Haidian District, 100094 Beijing, People’s Republic of China
X. Yang
State Key Laboratory for Agrobiotechnology, College of Biology, China Agricultural University, Yuanming Yuan West Road #2, Haidian District, 100094 Beijing, People’s Republic of China
X. An
State Key Laboratory for Agrobiotechnology, College of Biology, China Agricultural University, Yuanming Yuan West Road #2, Haidian District, 100094 Beijing, People’s Republic of China
Y. Chen
State Key Laboratory for Agrobiotechnology, College of Biology, China Agricultural University, Yuanming Yuan West Road #2, Haidian District, 100094 Beijing, People’s Republic of China
Myostatin (MSTN), a member of the transforming growth factor-β superfamily, has been shown to be a negative regulator of myogenesis. Natural mutation in beef cattle causes double-muscling phenotypes. We report an investigation designed to knockout the MSTN gene by gene targeting in ovine myoblast cells. Two promoter-trap targeting vectors MSTN-green fluorescent protein (GFP) and MSTN-neo were constructed and used to transfect foetal and neonatal ovine primary myoblast cells. Both GFP-expressing cells and drug-resistant cells were obtained. Targeted cells expressing GFP were confirmed by polymerase chain reaction (PCR) assay and drug-resistant cells were characterised by PCR and Southern blot after growing into cell clones.
