Karunakaran T, Gunasekaran P
Department of Microbiology and Microbial Technology, School of Biological Sciences, Madurai Kamaraj University, India.
Curr Microbiol. 1992 Nov;25(5):291-5. doi: 10.1007/BF01575864.
An alkaline phosphatase (phoA) gene from Zymomonas mobilis was isolated in Escherichia coli CC118 by use of the plasmid Bluescript KS+. The origin of the 6.4-kb DNA fragment in pZAP1 from the chromosome of Z. mobilis was confirmed by Southern blotting and hybridization studies. The Z. mobilis phoA gene was localized at one end of the chromosomal insert on plasmid pZAP1. The Z. mobilis phoA gene was expressed from its own promoter in E. coli, and the enzyme was localized to the periplasmic space. Z. mobilis alkaline phosphatase activity in E. coli was repressed in high-phosphate media and derepressed under a phosphate-limited growth condition. These results suggest that Z. mobilis alkaline phosphatase is subjected to normal regulation in E. coli.
利用质粒pBluescript KS +，在大肠杆菌CC118中分离出运动发酵单胞菌的碱性磷酸酶（phoA）基因。通过Southern印迹和杂交研究证实了pZAP1中6.4 kb DNA片段来自运动发酵单胞菌染色体。运动发酵单胞菌phoA基因位于质粒pZAP1上染色体插入片段的一端。运动发酵单胞菌phoA基因在大肠杆菌中由其自身启动子表达，且该酶定位于周质空间。在高磷酸盐培养基中，大肠杆菌中运动发酵单胞菌碱性磷酸酶活性受到抑制，而在磷酸盐限制的生长条件下则去抑制。这些结果表明，运动发酵单胞菌碱性磷酸酶在大肠杆菌中受到正常调控。
