小鼠Binlb基因的抗菌活性研究及其转基因小鼠的建立

STUDY ON THE ANTI-MICROBIAL ACTIVITY OF MOUSE BINIB AND CONSTRUCTION OF THE TRANSGENIC MICE WITH BIN1B OVER-EXPRESSION IN EPIDIDYMIS

我们在真核细胞中表达了小鼠Binlb基因,证实了它的抗菌活性,并进一步证明了该蛋白C端融合HA抗原决定簇后基本不影响蛋白的生物活性,为开展这一蛋白的转基因小鼠工作提供了较理想的转基因材料。同时我们利用四环素调控的基因表达系统(Tet-on系统)分别建立了附睾专一性表达rtTA的转基因小鼠和由TRE控制的mBinlb-HA的转基因阳性小鼠。这项研究对于我们理解mBinlb在体内的生理和病理作用提供了进一步研究的基础。

The cDNA of mouse Binlb homolog(mBinlb) protein was subcloned into expression vector of pCDNA3 and transiently expressed in COS-7 cells. The expression of mBinlb in COS-7 cells was identified by RT-PCR.The anti-microbial activity of the transfected cell culture supernatant was confirmed by plating on growth media. Adding a HA tag to the C-terminus of mBinlb (mBinlb-HA)resulted in the same anti-microbial activity as the mBinlb protein. This result indicated that anti-microbial activity was not located at the C terminus of mBinlb. To study the function of mBinlb in vivo, the Tet-on gene expression regulation system was used to construct transgenic mice. Two lines of transgenic mice were obtained, one in which the mBinlb-HA cDNA was under the control of the TRE promoter,and the other with rtTA specifically expressed using the promoter of the mE-RABP gene in the epididymis. Crossing the two transgenic lines will produce inducible over-expression of mBinlb specifically in epididymis. This research will provide a useful mice model to study the function of mBinlb in vivo.