关键词: 高原鼠兔, 瘦素蛋白, 乳腺上皮细胞, 乳腺特异表达载体, pBC1

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.

Key words: Leptin, Mammary epithelial cell, Mammary gland-specific expression vector, pBC1, Plateau pika