伤寒沙门菌pho基因缺陷变异株的制备_茅凌翔.pdf

DOI牶牨牥牣牨牫牫牨牪牤j牣issn牣牨牰牱牨牠牱牱牳牫牣牪牥牥牱牣牥牪牣牥牨牭 17 2 () Vo.l 17 No. 2 　 2007 3 Journal of Jiangsu University(M edicine Edition) 　 M ar. 2007 　 　 　 　 　 　 phoP 茅凌翔, 朱超望, 许化溪, 黄新祥 (, 212013) [　]　: phoP, phoP 。: phoP , P R, 5′Bg l II, P R phoP 、 , Bgl II, phoP 。 pG- MB151BamH I, , 。 P R, phoP , 。:P R, phoP 297。:phoP , 。 []　;phoP;; []　R392. 13　　[]　A　　[]　1671 -7783(2007)02 - 0145- 05 Construction of aphoP gene deleted mutant in Sa ml one lla en terica serovarTyphi MAO Ling-xiang, ZHU Chao-wang, XUHua-xi, HUANGX in-xiang (Department of Biochemistry, School ofM edical Technology, Jiangsu University, Zhenjiang Jiangsu 212013, hina) [Abstract]　Objective:For investigating functions of the regulator phoP ofSalmonella enterica serovar Typh,i phoP gene deletionmutant ofS. enterica serovar Typhiwas constructed. M ethods:A s the genom ic information, wt o pair′s prmi erswere designed at uppe-r and down-stream s of thephoP gene ofS. enterica serovar Typhi to amplify wt o homologousDNA fragments. ABg l II sitewas added in 5′term iniof the reverse prmi er of the uppe-r stream fragment and the forward prmi er of the down stream fragment. After amplification and purification, wt o fragmentswere digested byBgl II and ligated as the homologous recombinant DNA fragmen,t which contained the defective target gene. The homologous recombinant DNA fragmentwas in- serted into theBamH I site of the suicide plasm id pGMB151. The positive vectorwas then transferred into the target cell ofS. enterica serovar Typhiw ild strain by using of electroporation. The recombination was visualized by P R, and the complete recombinant strai