Cloning and structural analysis of the Klebsiella oxytoca VN13 peh gene

Abstract

Визначено иуклеотидпу послідовність промоторної області та 1450 п н. структурної частини гена полігалактуронази (peh) ендофітної бактерії К. oxytoca VN13, який, можливо, бере участь у механізмі проникнення бактерій у тканини коренів рослин. Досліджено рівень гомології ДНК та амінокислотної послідовності кодованого білка з відповідними послідовностями інших бактерій.

A PCR-mediated method for discrimination of Klebsiella oxytoca between closely related bacteria in environmental and clinical specimens

“…Amplification of the unique DNA fragments in the PCR test revealed high level of specificity of the primers designed for the pehX gene. The negative results in E. carotovora and E. chrysanthemi strains were expected because there was practically no homo logy between the polygalacturonase-encoding gene of K. oxytoca and analogous genes of Erwinia [21 ]. The absence of amplicons in K. pneumoniae, K. planticola and E. coli JM109 can be explained by the absence of specific gene encoding the hydrolytic enzyme.…”

“…Priming oligonuc leotides were synthesised at the Institute of Bioorganic Chemistry of RAN (Russian Federation). Results and Discussion Earlier the pehX gene of К oxytoca VN13 has been cloned and sequenced (accession No AY06648) [21 ]. The sequence was compared with the other peh gene sequences available in GenBank, and the comparison did not display essential homology in the DNA sequences of pectinolytic bacteria.…”

Molecular Discrimination of Klebsiella oxytoca using Polymerase Chain Reaction Targeted Polygalacturonase (pehX) Gene

“…This is a unique capability for K. oxytoca amongst the Klebsiella spp. If a sequence coding for this specific enzyme is amplified with PCR, proper discrimination of K. oxytoca from other Klebsiella strains can be achieved (Kovtunovych and Kozyrovska, 2000).…”