CLONING AND SEQUENCE-ANALYSIS OF THE GENE ENCODING 3-HEXULOSE-6-PHOSPHATE SYNTHASE FROM THE METHYLOTROPHIC BACTERIUM, METHYLOMONAS-AMINOFACIENS-77A, AND ITS EXPRESSION IN ESCHERICHIA-COLI

A DNA fragment of 550 bp was specifically amplified by PCR with primer s based on the N-terminal sequence of the purified 3-hexulose-6-phosph ate synthase from Methylomonas aminofaciens 77a and on that of a,lysyl endopeptidase-derived peptide. Using this PCR product as a probe, a g ene coding for 3-hexulose-6-phosphate synthase in M. aminofaciens 77a chromosomal DNA was cloned in Escherichia coli JM109. Sequencing analy sis revealed that the gene encoding 3-hexulose-6-phosphate synthase co ntained a 624-bp open reading frame, encoding a protein composed of 20 8 amino acid residues with a calculated relative molecular mass of 212 24.