转甲状腺素蛋白基因在酵母中的表达

用限制酶BamHI将含人TTR基因的DNA片段(约493bp)从质粒 pSK-TTR上切下后并插入到分泌型载体pHIL-SI的BamHI位点上，经酶切鉴定重组质粒pHIL-SI-TTR上的TTR基因插入方向正确。将pHIL-SI-TTRDNA用BglII酶切后，转入真核表达系统——毕赤氏酵母。得到的转化子经摇瓶发酵，上清液用15%SDSPAGE检测，有TTRF的表达。经DEAESepheroseF.F离子交换柱和SephacrylS200分子筛层析，得到纯化的TTRF。体外实验表明TTRF对肝癌细胞有抑制作用。

Expression of Transthyretin Gene in Yeast Pichia pastoris

Plasmid pSK TTR was digested by Bam HI and the DNA fragment containing TTR gene (Transthyretin) was cloned into the Bam HI site of secretion vector pHIL SI TTR.In right orientation and frame.After the recombinant plasmid pHIL SI TTR was digested by Bgl II,the larger fragment of it was transformed into yeast Pichia pastoris. The SDS PAGE analysis showed that the yeast transformants could express and secret TTRF.The pure TTRF could be obtained by DEAE Sepherose F.F.chromatography and SephacrylS 200 chromatography.The vitro test showed inhibition of the growth of hepatoma cells by TTRF obtained from our experiments.