Cloning sequence analysis and characterization of DNA polymerase from Geobacillus sp. WBI

Abstract

A thermostable DNA polymerase gene (DNA pol) was isolated from the genomic DNA of Geobacillus sp.WBI by polymerase chain reaction (PCR). PCR amplification with the primer pair designed from the conserved sequence of several Geobacillus spp. produced a single band of 1.9 kb DNA fragment. The PCR product was purified and sequenced. Nucleotide sequence analysis revealed a high degree of similarity with the corresponding sequences of thermophilic Bacillus adolyticus XM, Geobacillus thermoleovorans, Bacillus caldotenax, Geobacillus kaustophilus, Bacillus sp. G and Geobacillus sp. Y412MC52. The maximum similarity (99%) was shared by B. caldolyticus. Based on the sequence comparison, it was found that the organism contained a thermostable DNA pol in its genome. Since the thermostable DNA polymerase is extremely important biotechnologically, the present study encourages further research to fully characterize the enzyme and determine its potential applications.