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KMID : 0380219940270010013
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Journal of Biochemistry and Molecular Biology
1994 Volume.27 No. 1 p.13 ~ p.16
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Cloning and Expression of Lipase from Pseudomonas fragi as Fusion Protein in Escherichia
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Abstract
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Abstract:
@EN The gene coding for the triacylglycerol lipase (E.C. 3.1.1.3.) of Pseudomonas fragi IFO-12049 was amplified by polymearse chain reaction(PCR). The PCR was performed with two primers carrying EcoRI and HindIII sites, respectively land
chromosomal Dna
of P. Fragi was used as a template. The PCR product of 1050 base pairs(bp) holding the open-reading frame of lipase gene was ligated with EcoRIlinker and inserted into EcoRI site of t he pGEX-2T as expression vector. The fused gene was
transformed
into
E. coli mC1061 and amplified. The pGEX-2T harboring lipase gene was isolated and named pGL19. E. coli MC1061 carrying pGL19 was induced by isopropyl-β-D-thiogalactoside(IPTG) and the lipase was highly expressed in E coli as a fusion protein with
glutathione-S-transferase(GST) and exhibited strong lipase activity on the plate containing tributyrin.
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