Production GH transgenic goat improving mammogenesis by somatic cell nuclear transfer

• Published in: Molecular biology reports 2014-07, Vol.41 (7), p.4759-4768
• Main Authors: Zhang, Q, Chen, J. Q, Lin, J, Yu, Q. H, Yu, H. Q, Xu, X. J, Liu, G. H, Yang, Q
• Format: Article
• Language: English
• Subjects: Animal Anatomy; Animal Biochemistry; Animal lactation; animal models; Animals; Animals, Genetically Modified; Biomedical and Life Sciences; Blastomeres - cytology; Blastomeres - physiology; Cloning; Cloning, Organism; dairy goats; Electroporation; Embryo Transfer; enzyme-linked immunosorbent assay; epithelial cells; Female; fibroblasts; Fibroblasts - cytology; Fibroblasts - physiology; Gene Expression; gene overexpression; Genetic Vectors; Goats; Goats - genetics; Growth Hormone - genetics; Growth Hormone - metabolism; Growth hormones; Histology; lactation; Lactation - genetics; Life Sciences; Male; mammary development; mammary glands; Mammary Glands, Animal; milk production; Morphology; Nuclear Transfer Techniques; oocytes; Oocytes - cytology; Oocytes - physiology; Organ Size; phenotype; polymerase chain reaction; Pregnancy; progeny; somatic cells; somatotropin; Transgenes
• ISSN: 0301-4851; 1573-4978
• DOI: 10.1007/s11033-014-3347-7

Growth hormone is a positive regulator of mammary gland development. Dairy animals that are administered growth hormone display enhanced lactation performance, a desirable agricultural trait. The objective of the current research was to generate an improved milk production phenotype in a large animal model using over-expressed GH in the mammary gland to promote mammogenesis. To this end, we constructed a mammary gland-specific expression vector, pcGH, and demonstrated effective GH expression in goat mammary epithelial cells in vitro by ELISA. Then, to produce transgenic offspring that were capable of stable GH expression in vivo, the linearized pcGH vector was electroporated into goat fetal fibroblasts. Cell colonies that were positive for GH were used as donors for nuclear transfer to enucleated oocytes. A total of 253 morulae or blastocytes developed from the reconstructed embryos were transferred to 56 recipients, resulting in 24 pregnancies at day 35. Finally, six transgenic goats were born. PCR detection confirmed the success of the cloning procedure. To observe the mammogenesis of dairy goats, the GH transgenic goats were mated with a completely healthy buck. In the later pregnancy period, the mammary gland of the GH transgenic goats were extensive than non-transgenic goats. These experiments indicated that the pcGH vector was incorporated into the transgenic goats and affected mammogenesis, which laid a solid foundation for elucidating the impact of GH on mammogenesis and lactation performance.