Cloning and Characterization of Phosphomannomutase/Phosphoglucomutase (pmm/pgm) Gene of Vibrio anguillarum Related to Synthesis of LPS
2016
Oh, R., Pukyong National University, Busan, Republic of Korea | Moon, S.Y., Pukyong National University, Busan, Republic of Korea | Cho, H.J., Pukyong National University, Busan, Republic of Korea | Jang, W.J., Pukyong National University, Busan, Republic of Korea | Kim, J.H., Pukyong National University, Busan, Republic of Korea | Lee, J.M., Pukyong National University, Busan, Republic of Korea | Kong, I.S., Pukyong National University, Busan, Republic of Korea
The phosphomannomutase/phosphoglucomutase gene (pmm/pgm) of Vibrio anguillarum (the causative agent of fish vibriosis) was cloned, and the open reading frame corresponded to a protein with 446 amino acids. The pmm/pgm gene showed a significant degree of sequence homology with the previously reported genes from V. mimicus, V. vulnificus, V. splendidus, and V. harveyi, with 92.3%, 91.4%, 89.9%, and 89.9% amino acid identity, respectively. By reverse transcriptase-polymerase chain reaction, we found that the pmm/pgm gene was upregulated under cold stress condition. The PMM/PGM protein is known to catalyze the interconversion between mannose-1-phosphate and mannose-6-phosphate or glucose-1-phosphate and glucose-6-phosphate, which are important intermediates for lipopolysaccharide (LPS) biosynthesis. To confirm the role of PMM/PGM in the LPS biosynthetic pathway, we constructed a knock out mutant by homologous recombination. The respective LPSs were isolated from the V. anguillarum wild-type and mutant strains, and changes were compared by subjecting them to sodium dodecyl sulfate polyacrylamide gel electrophoresis. Based on the different patterns of the LPSs, we expect the pmm/pgm gene to have an important role in LPS biosynthesis. The pmm/pgm-deficient mutant of V. anguillarum will contribute to further studies about the role of LPS in V. anguillarum pathogenesis.
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