Isolation and Identification of Helicase from Anoxybacillus sp for DNA amplification

Abstract

Helicase-producing thermophilic microorganisms were successfully isolated from water samples collected from hot springs in the from Ulu Legong Hot spring, Kedah, Malaysia and were identified up to genus level. The light and scanning microscopy technique were used to identify the morphology of the isolate. Chromosomal DNA from the organism was isolated and used to amplify 16S rRNA and UvrD gene fragments. The gene was amplified by a set of universal primers (F_UNI16S and R_UNI16S). The phylogenetic tree, homological analysis, and detailed comparison of the sequences showed that 16S rRNA gene sequence of the isolate had closest similarities with Anoxybacillus sp. Isolates gave PCR fragments of 2149bp which represent the UvrD gene and 1500bp which represent the 16S rRNA respectively of Anoxybacillus. Anoxybacillus sp was successfully isolated and identified by using 16S rRNA gene sequence analysis with UvrD gene.