Myostatin gene targeting in cultured China Han ovine myoblast cells

• Main Authors: L. Zhang, X. Yang, X. An, Y. Chen
• Format: Article
• Language: English
• Published: Elsevier 2007-01-01
• Series: Animal
• Subjects: gene targeting; myoblast cell; myostatin transgenics
• Online Access: http://www.sciencedirect.com/science/article/pii/S1751731107000717

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.