A Study Of Establishment Of Transgenic Mice Sharing Inducible Expression Of Pgh

This study aims to explore the construction of eukaryotic expression vector carrying porcine growth hormone gene (pGH) by using molecular biology method. After transfected into PK15 cell lines, we have verified the induction efficiency and expression efficiency after adding Doxycycline (DOX) by water. The results were showed as following: pTRE-GH fragment was amplified by PCR from pTRE-GH12 vector, then the ligation of the Sal I digested fragment to the Sal I digest vector pCAGGS-rtTA, the recombinant vector pCAGGS-rtTA-TRE-GH12(pTTGH) was constructed. The pTTGH vector DNA transfected porcine PK15 cells, and enriched with G418, Quantitative fluorogenic real-time PCR (QRT-PCR) assay for pGH mRNA expression level after adding induction substrate, doxycycline (DOX) in culture medium. QRT-PCR and western blot assay for pGH after different concentration of DOX inducing. The results of sequencing and restriction showed that the pTTGH vector was constructed successfully which can realize pGH controllable expression. This study establishes a foundation for preparation pGH controllable expression transgenic animal and further study on pGH affection.