Rearing process for Shuangji cloned pig by gene engineering technology
A kind of genetic engineering and technical technology, applied in the field of "double muscle" cloned pigs, which can solve the problems of transgenic families that have not yet obtained homozygosity, and the characters cannot be stably inherited.
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[0014] Materials and methods
[0015] 1. Reagents and medium: All reagents were purchased from Sigma-Aldrich Company; endonucleases and molecular biology reagents used were purchased from Promega Company; medium was purchased from Gibco Company.
[0016] 2. Gene cloning and identification: The full-length sequence of the porcine myostatin gene was screened out from the porcine genome library by conventional methods, and sequence analysis was carried out [Stratil A, Kopecny M. Genomic organization, sequence and polymorphism of the porcine myostatin (GDF8; MSTN ) gene. Anim Genet, 30(6): 468-70.].
[0017] 3. Construction of porcine myostatin gene targeting vector. A vector for targeting is constructed by conventional molecular cloning methods using appropriate restriction sites. After successful digestion and confirmation, the vector was linearized and quantified for later use.
[0018] 4. Obtain mature oocytes from sexually mature or pre-sexually mature s...
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