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Improvement of cyclodextrin glycosyltransferase gene expression in Escherichia coli by insertion of regulatory sequences involved in the promotion of RNA transcription

Regulation of RNA transcription in controlling the expression of genes at promoter and terminator regions is crucial as the interaction of RNA polymerase occurred at both sites. Gene encoding cyclodextrin glycosyltransferase (CGTase) from Bacillus sp. NR5 UPM isolated in the previous study was used...

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Main Authors: Ramli, Norhayati, Abd. Aziz, Suraini, Mohammed Alitheen, Noorjahan Banu, Hassan, Mohd Ali, Maeda, Toshinari
Format: Article
Language:English
Published: Springer 2013
Online Access:http://psasir.upm.edu.my/id/eprint/28102/1/Improvement%20of%20cyclodextrin%20glycosyltransferase%20gene%20expression%20in%20Escherichia%20coli%20by%20insertion%20of%20regulatory%20sequences%20involved%20in%20the%20promotion%20of%20RNA%20transcription.pdf
http://psasir.upm.edu.my/id/eprint/28102/
http://link.springer.com/article/10.1007%2Fs12033-013-9647-7
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spelling my.upm.eprints.281022016-06-20T06:14:00Z http://psasir.upm.edu.my/id/eprint/28102/ Improvement of cyclodextrin glycosyltransferase gene expression in Escherichia coli by insertion of regulatory sequences involved in the promotion of RNA transcription Ramli, Norhayati Abd. Aziz, Suraini Mohammed Alitheen, Noorjahan Banu Hassan, Mohd Ali Maeda, Toshinari Regulation of RNA transcription in controlling the expression of genes at promoter and terminator regions is crucial as the interaction of RNA polymerase occurred at both sites. Gene encoding cyclodextrin glycosyltransferase (CGTase) from Bacillus sp. NR5 UPM isolated in the previous study was used for further construction of pTZCGT-SS, pTZCGT-BS and pTZCGT-BT expression systems for enhancement of CGTase production. The putative promoter regions, −35 and −10 sequences were found in the upstream of the mature gene start codon. Whereas, long inverted repeats sequences which can form a stable stem and loop structure was found downstream of the open reading frame (ORF) of Bacillus sp. NR5 UPM CGTase. The construction of E. coli strain harbouring pTZCGT-BS showed increment of 3.2-fold in CGTase activity compared to the wild type producer. However, insertion of terminator downstream of CGTase gene in E. coli strain harbouring pTZCGT-BT only resulted in 4.42 % increment of CGTase production compared to E. coli strain containing pTZCGT-BS, perhaps due to low intrinsic termination efficiency. Thus, it is suggested that the insertion of the putative promoter regions upstream of the coding sequence for the construction of CGTase expression system will further enhance in the recombinant enzyme production. Springer 2013 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/28102/1/Improvement%20of%20cyclodextrin%20glycosyltransferase%20gene%20expression%20in%20Escherichia%20coli%20by%20insertion%20of%20regulatory%20sequences%20involved%20in%20the%20promotion%20of%20RNA%20transcription.pdf Ramli, Norhayati and Abd. Aziz, Suraini and Mohammed Alitheen, Noorjahan Banu and Hassan, Mohd Ali and Maeda, Toshinari (2013) Improvement of cyclodextrin glycosyltransferase gene expression in Escherichia coli by insertion of regulatory sequences involved in the promotion of RNA transcription. Molecular Biotechnology, 54 (3). pp. 961-968. ISSN 1073-6085; ESSN: 1559-0305 http://link.springer.com/article/10.1007%2Fs12033-013-9647-7 10.1007/s12033-013-9647-7
institution Universiti Putra Malaysia
building UPM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Putra Malaysia
content_source UPM Institutional Repository
url_provider http://psasir.upm.edu.my/
language English
description Regulation of RNA transcription in controlling the expression of genes at promoter and terminator regions is crucial as the interaction of RNA polymerase occurred at both sites. Gene encoding cyclodextrin glycosyltransferase (CGTase) from Bacillus sp. NR5 UPM isolated in the previous study was used for further construction of pTZCGT-SS, pTZCGT-BS and pTZCGT-BT expression systems for enhancement of CGTase production. The putative promoter regions, −35 and −10 sequences were found in the upstream of the mature gene start codon. Whereas, long inverted repeats sequences which can form a stable stem and loop structure was found downstream of the open reading frame (ORF) of Bacillus sp. NR5 UPM CGTase. The construction of E. coli strain harbouring pTZCGT-BS showed increment of 3.2-fold in CGTase activity compared to the wild type producer. However, insertion of terminator downstream of CGTase gene in E. coli strain harbouring pTZCGT-BT only resulted in 4.42 % increment of CGTase production compared to E. coli strain containing pTZCGT-BS, perhaps due to low intrinsic termination efficiency. Thus, it is suggested that the insertion of the putative promoter regions upstream of the coding sequence for the construction of CGTase expression system will further enhance in the recombinant enzyme production.
format Article
author Ramli, Norhayati
Abd. Aziz, Suraini
Mohammed Alitheen, Noorjahan Banu
Hassan, Mohd Ali
Maeda, Toshinari
spellingShingle Ramli, Norhayati
Abd. Aziz, Suraini
Mohammed Alitheen, Noorjahan Banu
Hassan, Mohd Ali
Maeda, Toshinari
Improvement of cyclodextrin glycosyltransferase gene expression in Escherichia coli by insertion of regulatory sequences involved in the promotion of RNA transcription
author_facet Ramli, Norhayati
Abd. Aziz, Suraini
Mohammed Alitheen, Noorjahan Banu
Hassan, Mohd Ali
Maeda, Toshinari
author_sort Ramli, Norhayati
title Improvement of cyclodextrin glycosyltransferase gene expression in Escherichia coli by insertion of regulatory sequences involved in the promotion of RNA transcription
title_short Improvement of cyclodextrin glycosyltransferase gene expression in Escherichia coli by insertion of regulatory sequences involved in the promotion of RNA transcription
title_full Improvement of cyclodextrin glycosyltransferase gene expression in Escherichia coli by insertion of regulatory sequences involved in the promotion of RNA transcription
title_fullStr Improvement of cyclodextrin glycosyltransferase gene expression in Escherichia coli by insertion of regulatory sequences involved in the promotion of RNA transcription
title_full_unstemmed Improvement of cyclodextrin glycosyltransferase gene expression in Escherichia coli by insertion of regulatory sequences involved in the promotion of RNA transcription
title_sort improvement of cyclodextrin glycosyltransferase gene expression in escherichia coli by insertion of regulatory sequences involved in the promotion of rna transcription
publisher Springer
publishDate 2013
url http://psasir.upm.edu.my/id/eprint/28102/1/Improvement%20of%20cyclodextrin%20glycosyltransferase%20gene%20expression%20in%20Escherichia%20coli%20by%20insertion%20of%20regulatory%20sequences%20involved%20in%20the%20promotion%20of%20RNA%20transcription.pdf
http://psasir.upm.edu.my/id/eprint/28102/
http://link.springer.com/article/10.1007%2Fs12033-013-9647-7
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score 13.211869

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