%0 Journal Article
%T Expression of functional human STK11 protein in Escherichia coli<br>在大肠杆菌中表达有活性的人STK11蛋白
%A HOU Xin
%A HU Ting-mao
%A LIU Jun-e
%A <br>侯鑫
%A 扈廷茂
%A 刘俊娥
%J 微生物学报
%D 2007
%I 
%X STK11(serine/threonine kinase 11), a multi-functional protein reported recently, possibly participates in a broad range of cellular processes, including regulation of cell cycle, p53-mediated apoptosis, ras-induced cell transformation and cell polarization. An efficient expression of functional STK11 in Escherichia coli will promote the study on its structure and function. Inducible prokaryotic expression vector pET-Nus-STK11(with Nus fusion tag) was constructed with pET-44a( ) and the cDNA of STK11 gene cloned in our lab. pET-Nus-STK11 was then expressed in both BL21(DE3) and Rosetta-gami (DE3)pLysS on the induction of IPTG. SDS-PAGE and Western blot indicated that recombinant Nus-STK11 obtained in BL21(DE3) was in the form of inclusion body, whereas that from Rosetta-gami (DE3)pLysS was mainly in soluble fraction, and accounted for 8.9% and 16.7% of the total protein, respectively. After purification and refolding, the obtained recombinant protein was carried into SMMC-7721 cells by Chariot to observe its influence on cell growth and cell cycle. Nus-STK11 from BL21(DE3) was proved to be lack of any tumor-suppression activity, while a growth inhibitory ratio of 47.05% on SMMC-7721 cell was observed, and cell cycle progression of SMMC-7721 cells was also arrested from G_0/G_1 to S phase, with the Nus-STK11 from Rosetta-gami (DE3)pLysS, indicating that the above recombinant fusion protein from Rosetta-gami (DE3)pLysS had significant biological activity. This is the first report on functional recombinant STK11 protein expressed in Escherichia coli.
%K Rosetta-gami(DE3)pLysS<br>STK11蛋白
%K 原核表达
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A3C6BA55AB623B90FA9104CFFC826F3C&aid=28214D8522FBAB6B&yid=A732AF04DDA03BB3&vid=F4B561950EE1D31A&iid=CA4FD0336C81A37A&sid=9C65ADEB5990B252&eid=0D0D661F0B316AD5&journal_id=0001-6209&journal_name=微生物学报&referenced_num=0&reference_num=16