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Establishment of MSTN Interfering Plasmid Transgenic Cell Lines and Integration of Exogenous Gene
Liu Dan, Tong Huili, Li Shufeng, Chang Shuwei, Yang Cuicui, Yan Yunqin*
College of Life Science, Northeast Agricultural University, Harbin 150030, China
Abstract: In this study， we used MSTN interference vector to transfect Luxi cattle fetal fibroblasts and obtained five transgenic monoclonal cell lines. By Real-time PCR and high-efficiency thermal asymmetric interlaced PCR， we detected the expressing copies of MSTN plasmid and their integration sites in bovine genome. The results showed that the copy numbers of plasmid were effectively detected by quantitative PCR in the five positive cell clones which were 2.26±0.32， 1.52±0.25， 25.68±1.02， 8.43±0.73 and 6.72±0.1， respectively. Integration sites detected by hiTAIL-PCR showed that there was recombination in the process of insertion of plasmid fragment into target genome. This study explored the integration mechanism of MSTN carrier interference in bovine fetal fibroblasts， in order to obtain the clear genetic background of transgenic cells. It is important for obtaining transfer material by somatic cell nuclear， providing important theoretical and experimental basis for cultivation of highyielding new varieties of transgenic cattle.
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