01Jan 2017

Molecular cloning of putative L-asparaginase/glutaminase gene from Klebseilla peunomonae MGH 78578

Abstract


This study is one among a few local studies that have been reported to clone the putative L-asparaginase/glutaminase produced by recombinant gene based on the use of plasmid Ecoli - Nterm 6X HN(PLEICS-01) vector. In this study, a pair of primers was designed to obtain L- glutamin-(asparagine)ase encoding gene from Klebsiella pneumoniae subsp. pneumoniae MGH 78578. The putative L-asparaginase/glutaminase gene encoding gene (KPN-01165) was expressed in E.coli BL21(D3) to make sure the presence of a functional putative L-asparaginase/glutaminase in the cloned DNA. The recombinant L- glutamin-(asparagine)ase gene was 1005 bps which encoding a protein of 334 amino acids. The recombinant enzyme containing a 6xHistidine-tag can easily purified by Ni-NTA chromatography and its molecular weight was determined to be 40 KDa by SDS-PAGE. A putative L-asparaginase/glutaminase catalysed the hydrolysis is not only of L-glutamine, but also of L-asparagine.


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[Maha Hameed, Abdul kareem A.Al Kazaz. and Ghazi M.Aziz (2014); Molecular cloning of putative L-asparaginase/glutaminase gene from Klebseilla peunomonae MGH 78578 Int. J. of Adv. Res. 2 (6). 0] (ISSN 2320-5407). www.journalijar.com


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