Heterologous Expression Of Violacein Biosynthetic Gene Cluster From Chromobacterium Violaceum Strain Mth01 In Escherichia Coli

Violacein, a bisindole pigment produced by Chromobacterium violaceum has tremendous pharmaceutical potential such as antimicrobial, antiparasites and antiviral. The violacein structure is dimeric composed of three structural units, a 5- hydroxyindole, oxindole and 2- pyyrolidone subunits formed b...

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主要作者: Matthew Minggu, Matthlessa
格式: Thesis
语言:English
出版: 2017
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spelling my-usm-ep.609302024-08-13T08:50:08Z Heterologous Expression Of Violacein Biosynthetic Gene Cluster From Chromobacterium Violaceum Strain Mth01 In Escherichia Coli 2017-03 Matthew Minggu, Matthlessa RM260-263 Chemotherapy Violacein, a bisindole pigment produced by Chromobacterium violaceum has tremendous pharmaceutical potential such as antimicrobial, antiparasites and antiviral. The violacein structure is dimeric composed of three structural units, a 5- hydroxyindole, oxindole and 2- pyyrolidone subunits formed by the condensation of two modified tryptophan molecules. Violacein is made by a series of five specific genes termed vzoA, vzoB, vioC, vioE), and vzoE, giving its total sequence length of 7.3 kilo-bases (kb) in size. The present work describes the development of a genetic system for heterologous production of violacein in Escherichia coli cells. C. violaceum strain MTH01, a natural violacein producing bacterium was isolated from the limnological areas of Tasik Harapan at Universiti Sains Malaysia, Pulau Pinang. The complete violacein biosynthetic gene cluster was successfully amplified from MTH01 genomic DNA using violacein gene specific primers, and cloned into pUC19 vector. Sequence analysis of the DNA insert harboured by the recombinant pUC19 clone, pVio!9, showed that the cloned fragment shared 99% homology to published sequences of the C. violaceum violacein biosynthetic gene cluster. The sequenced fragment was subcloned into pET-3a and pET-1 lb expression vectors, yielding recombinant vectors, pVio3a and pViollb, respectively. 2017-03 Thesis http://eprints.usm.my/60930/ http://eprints.usm.my/60930/1/interologous%20expression%20of%20violacein%20cut.pdf application/pdf en public masters Universiti Sains Malaysia Pusat Pengajian Sains Farmasi
institution Universiti Sains Malaysia
collection USM Institutional Repository
language English
topic RM260-263 Chemotherapy
spellingShingle RM260-263 Chemotherapy
Matthew Minggu, Matthlessa
Heterologous Expression Of Violacein Biosynthetic Gene Cluster From Chromobacterium Violaceum Strain Mth01 In Escherichia Coli
description Violacein, a bisindole pigment produced by Chromobacterium violaceum has tremendous pharmaceutical potential such as antimicrobial, antiparasites and antiviral. The violacein structure is dimeric composed of three structural units, a 5- hydroxyindole, oxindole and 2- pyyrolidone subunits formed by the condensation of two modified tryptophan molecules. Violacein is made by a series of five specific genes termed vzoA, vzoB, vioC, vioE), and vzoE, giving its total sequence length of 7.3 kilo-bases (kb) in size. The present work describes the development of a genetic system for heterologous production of violacein in Escherichia coli cells. C. violaceum strain MTH01, a natural violacein producing bacterium was isolated from the limnological areas of Tasik Harapan at Universiti Sains Malaysia, Pulau Pinang. The complete violacein biosynthetic gene cluster was successfully amplified from MTH01 genomic DNA using violacein gene specific primers, and cloned into pUC19 vector. Sequence analysis of the DNA insert harboured by the recombinant pUC19 clone, pVio!9, showed that the cloned fragment shared 99% homology to published sequences of the C. violaceum violacein biosynthetic gene cluster. The sequenced fragment was subcloned into pET-3a and pET-1 lb expression vectors, yielding recombinant vectors, pVio3a and pViollb, respectively.
format Thesis
qualification_level Master's degree
author Matthew Minggu, Matthlessa
author_facet Matthew Minggu, Matthlessa
author_sort Matthew Minggu, Matthlessa
title Heterologous Expression Of Violacein Biosynthetic Gene Cluster From Chromobacterium Violaceum Strain Mth01 In Escherichia Coli
title_short Heterologous Expression Of Violacein Biosynthetic Gene Cluster From Chromobacterium Violaceum Strain Mth01 In Escherichia Coli
title_full Heterologous Expression Of Violacein Biosynthetic Gene Cluster From Chromobacterium Violaceum Strain Mth01 In Escherichia Coli
title_fullStr Heterologous Expression Of Violacein Biosynthetic Gene Cluster From Chromobacterium Violaceum Strain Mth01 In Escherichia Coli
title_full_unstemmed Heterologous Expression Of Violacein Biosynthetic Gene Cluster From Chromobacterium Violaceum Strain Mth01 In Escherichia Coli
title_sort heterologous expression of violacein biosynthetic gene cluster from chromobacterium violaceum strain mth01 in escherichia coli
granting_institution Universiti Sains Malaysia
granting_department Pusat Pengajian Sains Farmasi
publishDate 2017
url http://eprints.usm.my/60930/1/interologous%20expression%20of%20violacein%20cut.pdf
_version_ 1811772863400640512