Purification, gene cloning, amino acid sequence analysis, and expression of an extracellular lipase from an Aeromonas hydrophila human isolate

Article Abstract:

Lipase purified from both Aeromonas hydrophila culture supernatant and the periplasmic fluids of Escherichia coli comprising the lip determinant in the original clone (plasmid pLA2) indiciated an M(r) of 67,000 on sodium dodecyl sulfatepolyacrylamide gel electrophoresis which is consistent with the M(r) evaluated by Sephacryl S-200 chromatography. The cloning in Escherichia coli was done using pBR322 as a vector. Sequence analysis indicated a major open reading frame of 2,052 bp, predicting polypeptide with an M(r) of 71,804. The polypeptide comprised an amino acid sequence (V-H-F-L-G-H-S-L-G-A) among lipases.

Analysis, Escherichia coli, Lipase

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Molecular cloning of the tryptophan operon from an Aeromonas hydrophila freshwater isolate

Article Abstract:

A molecular genetic analysis of the tryptophan biosynthetic genes in Aeromonas hydrophila was undertaken. Complementation analysis of an A. hydrophila genomic library showed the presence of trpA, trpB, trpC, trpD and trpE genes. Furthermore, the analysis suggested that these genes were organized in a single operon. An internal secondary promoter in the distal portion of trpD was detected, suggesting that A. hydrophila is more closely related to the Enterobacteriaceae than to Vibrionaceae.

Genetic aspects, Operons, Anaerobic bacteria, Tryptophan metabolism

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Identification of Aeromonas hydrophila hybridization group 1 by PCR assays

Article Abstract:

A PCR assay using synthetic oligonucleotide primers of 23 and 24 bases amplifies a sequence of the lip gene that codes for the extracellular lipase of Aeromonas hydrophila. A 760 bp gene fragment was amplified from lysed A. hydrophila, which is pathogenic in humans. The amplified sequence can be identified using ethidium bromide-stained agarose gels or Southern blot analysis. PCR assays using cells of 50 strains of bacteria establish the primer specificity for A. hydrophila.

Usage, Polymerase chain reaction, Bacteria, Pathogenic, Pathogenic bacteria, Gene amplification

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