Isolation and Identification of Helicase from Escherichia coli for Biotechnology Processes

Abstract

Helicases are able to unwind duplex DNA enzymatically at one temperature without prior heat denaturation. Therefore, the current study was undertaken to isolate and identify E. coli for in house production of helicase enzymes. Bacteria were isolated from patient stool samples from Penang General Hospital, Penang, Malaysia. 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 E. coli. Isolates gave PCR fragments of 2163bp which represent the uvrD gene and 1500bp which represent the 16S rRNA respectively of E. coli. E. coli was successfully isolated and identified by using 16S rRNA gene sequence analysis with uvrD gene