Isolation And Identification Of UvrD Helicase From Escherichia Coli From Local Fever Patient For PCR Enzyme Production | 4864
Journal of Biotechnology & Biomaterials
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Helicase is an enzyme that plays essential roles in nearly all DNA metabolic transactions and has been implicated in a variety
of human genetic disorders. UvrD is a highly conserved helicase involved in DNA repair and it plays a critical role in
maintaining genomic stability and facilitating DNA lesion repair in many prokaryotic species as it also unwinds and separates
the two strands of the DNA double helix. Helicases are essential during DNA replication because they separate double-stranded
DNA into single strands allowing each strand to be copied. The objective of this study is to isolate and identify Helicase of local
E. coli isolate from fever patient. Nutrient broth and Nutrient agar was used for the isolation of the pathogenic microorganism
from the patient samples by using the streak plate method. The morphology properties of the microorganism were investigated
by using light microscope by Gram staining to confirm Gram reaction. Light micrograph and scanning electron microscopy
were performed to further identify the ultrastructure of the bacteria. Chromosomal DNA from the organism was isolated and
used to amplify 16S rRNA and uvrD gene fragments to detect the presence of helicase gene in the organisms. The clinical isolate
was grown well on MacConkey agar. The isolates are Gram-negative and rod in shape. The fact that the E. coli sample contains
the helicase gene was further supported by the presence of UvrD gene from the gel electrophoresis image. In conclusion, based
on the results of this study, Helicase (UvrD gene) was successfully isolated from fever patient infected with E. coli for various
biotechnological processes. This study will lead to in house production of helicase which could be applied in low setting diagnostic
kit development for third world country.
Yamuna Sundram has completed Bsc in Biotechnology from Asian Institute of Medicine, Science and Technology University and currently pursuing
MSc in Molecular Medicine at Institute for Research in Molecular Medicine, University Science Malaysia. This project funded by University Science
Malaysia research grant RUC (1001/ PSKBP/86300110).
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