Open AccessCloning 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
Author(s) -
Yanase Hideshi,
Ikeyama Kohei,
Mitsui Ryoji,
Ra Sonmin,
Kita Keiko,
Sakai Yasuyoshi,
Kato Nobuo
Publication year - 1996
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1996.tb07990.x
Subject(s) - escherichia coli , microbiology and biotechnology , atp synthase , gene , biology , peptide sequence , nucleic acid sequence , molecular cloning , open reading frame , sequence analysis , biochemistry , genetics
Abstract A DNA fragment of 550 bp was specifically amplified by PCR with primers based on the N‐terminal sequence of the purified 3‐hexulose‐6‐phosphate synthase from Methylomonas aminofaciens 77a and on that of a lysyl endopep(idase‐derived peptide. Using this PCR product as a probe, a gene coding for 3‐hexulose‐6‐phosphate synthase in M. aminofaciens 77a chromosomal DNA was cloned in Escherichia coli JM109. Sequencing analysis revealed that the gene encoding 3‐hexulose‐6‐phosphate synthase contained a 624‐bp open reading frame, encoding a protein composed of 208 amino acid residues with a calculated relative molecular mass of 21 224.