%0 Journal Article
%T Cloning,expression and characterization of a new hybrid AMP gene of Hex-Mag<br>一种新融合抗菌肽Hex-Mag基因的克隆、表达及其抗菌活性的研究
%A LI Gui-ping
%A CHEN Yi-ben
%A <br>李桂平
%A 陈仪本
%J 微生物学报
%D 2007
%I 
%X To enhance the antibacterial ability of Magainin1-12,its N side was joined with an alkaline peptide named Hexapeptide(RRWQWR),which would make Magainin1-12 cling to the membrane of bacterial cells even tighter. According to the partiality codon of Pichia pastoris,a new hybrid antibacterial peptide Hex-Mag was designed based on the sequence of Hexapeptide and Magainin(1-12). Synthesized through gene splicing by overlap extension,the hybrid gene was cloned into pPIC9 to construct the expression vector pPIC9-HM. After restriction enzyme analysis and purification, the pPIC9-HM was transformed into Pichia pastoris GS115. And the positive clones screened by the phenotype were induced by methanol. After optimized the requirements for the flask-shaking culture fermentation,the hybrid antibacterial peptide was expressed on high level. The new peptide,which has a weight of 2.3kDa,could remain its inhibition activity after treating for more than 3 hours in boiled water. Detected by agrose diffusion assay,Hex-Mag showed its broad-spectrum antibacterial abilities not only to Gram-negative bacteria but also to Gram-positive bacteria. The function of additive positive charges were testified by the antibacterial experiments, and the results showed the activity of Hex-Mag was stronger than that of Magainin1-12 obviously.
%K Hexapeptide(RRWQWR)
%K Magainin1-12<br>重叠区扩增基因拼接法
%K 融合肽Hex-Mag
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A3C6BA55AB623B90FA9104CFFC826F3C&aid=100CE9EA8A901958&yid=A732AF04DDA03BB3&vid=F4B561950EE1D31A&iid=CA4FD0336C81A37A&sid=EDA22B444205D04A&eid=2B5DE8A23DCEED39&journal_id=0001-6209&journal_name=微生物学报&referenced_num=0&reference_num=18