Abstract Ksta 2019.docx

“CLONING AND EXPRESSION OF GLUTATHIONE S-TRANSFERASE GENE IN PROKARYOTIC SYSTEM” Mamatha J1* ., Ishfaq Hassan., MirKhan .,Mohd Rijvan., Milton Mali.,Saikh Parvez & Chetana S *Email correspondence: [email protected] 1* PG -Department of Biotechnology, The Oxford College of Science, (TOCS) Bangalore. Glutathione S-Transferases (GST) represent a complex grouping of proteins widely distributed in prokaryotes and are grouped into several classes. Bacterial GSTs are implicated in a variety of distinct processes such as the biodegradation of xenobiotics, protection against chemical and oxidative stresses and antimicrobial drug resistance and also involved in a variety of distinct metabolic processes such as the biotransformation of dichloromethane, the degradation of lignin and atrazine. In present study aimed at cloning and expression of GST gene in E.coli K-12 substr. MG1655 . Genomic DNA from Escherichia coli K-12 substr. MG1655 were isolated and purified using agarose gel electrophoresis. The concentration of DNA was determined using a spectrophotometer at 260/280 nm. Primers for GST gene were designed by Oligo Calc and NEB cutter software and its amplification by PCR techniques. BamH1 and Xho1 restriction endonucleases were used to digest both pET-32b (+) vector and GST gene . Ligation of GST B gene into pET-32b (+) vector were successful with the help of T4 DNA Ligase followed by transformation of ligated GST B gene in to Prokaryotic system. After transformation the plates were observed for analyzing transformation efficiency. Two separate colonies were visualized on the agar plate. White coloured colonies contained the gene of interest and thus were transformed, whereas the blue colonies were deficit of the gene of interest and showed no transformation. Specific activity for E.coli was found to be 31.25 µmole/ml/min. Key words: Glutathione S-Transferases, E.coli, pET-32b(+) Vector, DNA Ligase.

Corresponding author address: Mamatha J. Associate Professor, P.G. Department of Biotechnology, The Oxford College of Science, Bangalore- 560 102, Karnataka, India Tel: +91 9738109917, Email: [email protected]