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Myostatin gene targeting in cultured China Han ovine myoblast cells

Published online by Cambridge University Press:  01 November 2007

L. Zhang ,
X. Yang ,
X. An  and
Y. Chen
L. Zhang
Affiliation:
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*
Affiliation:
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*
Affiliation:
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
Affiliation:
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
*
E-mail: [email protected]
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Abstract

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.

Keywords

gene targetingmyoblast cellmyostatin transgenics
Type
Full Paper
Information
animal , Volume 1 , Issue 10 , November 2007 , pp. 1401 - 1408
Copyright
Copyright © The Animal Consortium 2007

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