Construction of mammary gland-specific expression vector for plateau pika(Ochotona curzoniae)Leptin gene and its expression in cells

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  • (1 Key Laboratory of Adaptation and Evolution of Plateau Biota, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining 810001, Qinghai, China)
    (2 University of Chinese Academy of Sciences, Beijing 100049, China)
    (3 College of Biotechnology, Guilin Medical University, Guilin 541004, Guangxi, China)
    (4 College of Eco-Environment & Resources, Qinghai University For Nationalities, Xining 810007, Qinghai, China)
    (5 Molecular Pharmacology Research Center, School of Pharmaceutical Science, Wenzhou Medical University, Wenzhou 325035, Zhejiang, China)
    (6 College of Animal Science and Technology, Hunan Agricultural University, Changsha 410128, Hunan, China)

Online published: 2019-01-18

Abstract

Leptin is one of the key factors regulating body energy homeostasis. Previous research showed that the plateau pika(Ochotona curzoniae)leptin gene has undergone adaptive evolution. Functional experiments revealed that pika leptin regulates energy homeostasis via reduced food intake and increased energy expenditure under both warm and cold conditions. Together, these findings suggest that leptin of the plateau pika has a potentially potent role in regulating adaptive thermogenesis. In this study, the 504bp open-reading frame of obese(ob)gene was amplified from the full-length pika leptin cDNA and then cloned into a mammary gland-specific expression vect- or pBC1(pBC1-lep). Meanwhile, we successfully established dairy goat mammary epithelial cell lines by primary tissue mass cell culture and transfected the modified pBC1-lep plasmid into the cultured cells with lipofectamine, and then screened for and successfully established positive cell lines transfected with leptin. These results provide a probable way for expression of the plateau pika leptin in goat mammary gland specifically by transgenic animal and complete the construction of the eukaryotic mammary gland-specific expression vector.

Cite this article

PANG Bo, YU Honghao, ZHANG Xuze, XIE Ling, FU Lin, ZHAO Xinquan, GUO Songchang . Construction of mammary gland-specific expression vector for plateau pika(Ochotona curzoniae)Leptin gene and its expression in cells[J]. ACTA THERIOLOGICA SINICA, 2019 , 39(1) : 84 -93 . DOI: 10.16829/j.slxb.150172

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