Molecular cloning of the glyceraldehyde-3-phosphate dehydrogenase genes of Bacillus stearothermophilus and Escherichia coli, and their expression in Escherichia coli

Abstract

Recombinant plasmids derived from pBR322, which carry gap genes coding for the D-glyceraldehyde-3-phosphate dehydrogenases (GAPDH) of Bacillus stearothermophilus and Escherichia coli, have been isolated. The selection was carried out by complementation of an E. coli gapam mutant. Two plasmids containing B. stearothermophilus and E. coli DNA inserts of 4.3 kb and 1.4 kb, respectively, were characterized. Transformation of the E. coli mutant with either of the recombinant plasmids lead to a very high expression of the GAPDH activity. GAPDH produced by the strain containing the B. stearothermophilus gap gene was characterized by immunological cross-reactivity with antiserum raised against pure B. stearothermophilus GAPDH.