Matthew Minggu, Matthlessa
(2017)
Heterologous Expression Of
Violacein Biosynthetic Gene Cluster
From Chromobacterium Violaceum Strain
Mth01 In Escherichia Coli.
Masters thesis, Universiti Sains Malaysia.
Abstract
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.
Actions (login required)
 |
View Item |
