Construction of Targeting Vector for Swine MSTN Gene Knockout

摘要

MSTN gene is a negatively regulating factor of skeletal muscle growth and differentiation. This gene expression can be detected in all skeletal muscles of adult animals, mutation or deletion can increase the amount of muscles. The primers were designed based on the known sequence and homologous arms were obtained by PCR, homologous long arm is 5382bp, including the whole exon1 and exon2, homologous short arm is 844bp, including partial exon3, and constructs the targeting vector. The vector was designed with positive and negative selection strategy with insertion of homologous arms, and constructed by common molecular clogging techniques. Structure of this vector was identified by PCR and restriction analysis. The construction of targeting vector provides the experimental base for getting the MSTN gene deletion cell line.