运动发酵单胞菌中编码一种对磷酸盐不敏感的碱性磷酸酶的phoA基因在大肠杆菌中的克隆与表达。

Cloning and expression in Escherichia coli of a phoA gene encoding a phosphate-irrepressible alkaline phosphatase of Zymomonas mobilis.

作者信息

Michel G P, Alvarez E, Guzzo J, Cami B, Baratti J

机构信息

Centre National de la Recherche Scientifique, Laboratorie de Chimie Bactérienne, Marseille, France.

出版信息

FEMS Microbiol Lett. 1992 Nov 1;77(1-3):103-8. doi: 10.1016/0378-1097(92)90139-f.

DOI:10.1016/0378-1097(92)90139-f

PMID:1459397

Abstract

The Zymomonas mobilis phoA gene, encoding a phosphate-irrepressible alkaline phosphatase (ZAPase), was cloned and its expression was studied in phoA mutants of Escherichia coli. The ZAPase was recovered in the soluble fraction of E. coli. The enzyme was synthesized constitutively and its synthesis not repressed by phosphate, unlike the phoA gene of E. coli. The phoA gene of Z. mobilis was mutagenized by Mini Mu PR13 and the mutated gene crossed into Z. mobilis in order to obtain phoA mutants by reverse genetics. Although Z. mobilis mutants with Mini Mu PR13 integrated in the chromosome were obtained, none had an allele replacement for none was defective in ZAPase.

摘要

运动发酵单胞菌的phoA基因编码一种不受磷酸盐抑制的碱性磷酸酶（ZAPase），该基因被克隆并在大肠杆菌的phoA突变体中研究其表达。ZAPase在大肠杆菌的可溶部分中回收。该酶组成型合成，与大肠杆菌的phoA基因不同，其合成不受磷酸盐抑制。运动发酵单胞菌的phoA基因通过Mini Mu PR13诱变，诱变后的基因导入运动发酵单胞菌，以便通过反向遗传学获得phoA突变体。虽然获得了染色体中整合有Mini Mu PR13的运动发酵单胞菌突变体，但没有一个发生等位基因替换，因为没有一个ZAPase有缺陷。