Construction and function of mammary gland specific goat GH expression vector.
The objective of this study was to construct a mammary gland specific gene targeting vector (pLG), which could help the exogenous goat growth hormone (GH) cDNA being integrated into the beta-casein locus of goats and improve the milk production of goat. We cloned the 5' and 3' regulatory sequences of goat's beta-casein gene as homologous arms and inserted them into the Not I/Xho I and Sal I/Cla I multiple clone sites (MCS) of pLoxp II vector respectively, then inserted the GH cDNA into the Xho I MCS of pLoxp II vector and finally got the gene targeting vector pLG. The expression of GH in Bcap-37 cells transfected with plasmid pLG (14.52 ng/mL) was two times higher than that of cells untreated (7.10 ng/mL). While the GH expression of goats injected with plasmid pLG (7.54 ng/mL) was higher than goats untreated or injected with normal saline (6.64 and 6.78 ng/mL). We successfully constructed the mammary gland specific beta-casein gene targeting vector pLG exhibiting bioactivity of transcription and expression of GH both in vitro and in vivo.
Veterinary College, Nanjing Agricultural University, Weigang 1, Nanjing 210095, Jiangsu, People's Republic of China.
Qinghua
Q
Yu
Qi
Q
Tian
Jian
J
Lin
Qiang
Q
Zhang
Liqi
L
Zhu
Qian
Q
Yang
D000818:Animals; D002364:Caseins / Q000235:genetics; D005260:Female; D005347:Fibroblasts; D015870:Gene Expression; D023061:Gene Order; D018390:Gene Targeting; D056426:Genetic Loci; D005822:Genetic Vectors; D006041:Goats; D013006:Growth Hormone / Q000235:genetics* / Q000378:metabolism; D059765:Homologous Recombination; D008321:Mammary Glands, Animal / Q000378:metabolism*; D014158:Transcription, Genetic
D002364:Caseins; D013006:Growth Hormone