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Cloning and characterisation of the glyceraldehyde 3-phosphate dehydrogenase gene of Candida bombicola and use of its promoter

Inge Van Bogaert UGent, Sofie De Maeseneire UGent, Dirk Develter, Wim Soetaert UGent and Erick Vandamme UGent (2008) JOURNAL OF INDUSTRIAL MICROBIOLOGY AND BIOTECHNOLOGY. 35(10). p.1085-1092
abstract
The glyceraldehyde-3-phosphate dehydrogenase gene (GPD) of the sophorolipid producing yeast Candida bombicola was isolated using degenerated PCR and genome walking. The obtained 3,740 bp contain the 1,008 bases of the coding sequence and 1,613 and 783 bp of the upstream and downstream regions, respectively. The corresponding protein shows high homology to the other known GPD genes and is 74% identical to the gyceraldehyde-3-phosphate dehydrogenase of Yarrowia lipolytica. The particular interest in the C. bombicola GPD gene sequence originates from the potential use of its promoter for high and constitutive expression of homologous and heterologous genes. Southern blot analysis did not give any indication for the presence of multiple GPD genes and it can therefore be expected that the promoter can be used for efficient and high expression. This hypothesis was further confirmed by the biased codon usage in the GPD gene. GDP promoter fragments of different lengths were used to construct hygromycin resistance cassettes. The constructs were used for the transformation of C. bombicola and all of them, even the ones with only 190 bp of the GPD promoter, were able to render the cells resistant to hygromycin. The efficacy of a short GPD promoter can be a convenient characteristic for the construction of compact expression cassettes or vectors for C. bombicola. The GenBank accession number of the sequence described in this article is EU315245.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
sophorolipids, Candida bombicola, glyceraldehyde 3-phosphate dehydrogenase, promoter, hygromycin resistance
journal title
JOURNAL OF INDUSTRIAL MICROBIOLOGY AND BIOTECHNOLOGY
J. Ind. Microbiol. Biotechnol.
volume
35
issue
10
pages
1085 - 1092
Web of Science type
Article
Web of Science id
000259142400002
JCR category
BIOTECHNOLOGY & APPLIED MICROBIOLOGY
JCR impact factor
1.919 (2008)
JCR rank
74/144 (2008)
JCR quartile
3 (2008)
ISSN
1367-5435
DOI
10.1007/s10295-008-0386-x
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
524714
handle
http://hdl.handle.net/1854/LU-524714
date created
2009-03-19 09:38:21
date last changed
2009-04-01 13:43:14
@article{524714,
  abstract     = {The glyceraldehyde-3-phosphate dehydrogenase gene (GPD) of the sophorolipid producing yeast Candida bombicola was isolated using degenerated PCR and genome walking. The obtained 3,740 bp contain the 1,008 bases of the coding sequence and 1,613 and 783 bp of the upstream and downstream regions, respectively. The corresponding protein shows high homology to the other known GPD genes and is 74\% identical to the gyceraldehyde-3-phosphate dehydrogenase of Yarrowia lipolytica. The particular interest in the C. bombicola GPD gene sequence originates from the potential use of its promoter for high and constitutive expression of homologous and heterologous genes. Southern blot analysis did not give any indication for the presence of multiple GPD genes and it can therefore be expected that the promoter can be used for efficient and high expression. This hypothesis was further confirmed by the biased codon usage in the GPD gene. GDP promoter fragments of different lengths were used to construct hygromycin resistance cassettes. The constructs were used for the transformation of C. bombicola and all of them, even the ones with only 190 bp of the GPD promoter, were able to render the cells resistant to hygromycin. The efficacy of a short GPD promoter can be a convenient characteristic for the construction of compact expression cassettes or vectors for C. bombicola. The GenBank accession number of the sequence described in this article is EU315245.},
  author       = {Van Bogaert, Inge and De Maeseneire, Sofie and Develter, Dirk and Soetaert, Wim and Vandamme, Erick},
  issn         = {1367-5435},
  journal      = {JOURNAL OF INDUSTRIAL MICROBIOLOGY AND BIOTECHNOLOGY},
  keyword      = {sophorolipids,Candida bombicola,glyceraldehyde 3-phosphate dehydrogenase,promoter,hygromycin resistance},
  language     = {eng},
  number       = {10},
  pages        = {1085--1092},
  title        = {Cloning and characterisation of the glyceraldehyde 3-phosphate dehydrogenase gene of Candida bombicola and use of its promoter},
  url          = {http://dx.doi.org/10.1007/s10295-008-0386-x},
  volume       = {35},
  year         = {2008},
}

Chicago
Van Bogaert, Inge, Sofie De Maeseneire, Dirk Develter, Wim Soetaert, and Erick Vandamme. 2008. “Cloning and Characterisation of the Glyceraldehyde 3-phosphate Dehydrogenase Gene of Candida Bombicola and Use of Its Promoter.” Journal of Industrial Microbiology and Biotechnology 35 (10): 1085–1092.
APA
Van Bogaert, Inge, De Maeseneire, S., Develter, D., Soetaert, W., & Vandamme, E. (2008). Cloning and characterisation of the glyceraldehyde 3-phosphate dehydrogenase gene of Candida bombicola and use of its promoter. JOURNAL OF INDUSTRIAL MICROBIOLOGY AND BIOTECHNOLOGY, 35(10), 1085–1092.
Vancouver
1.
Van Bogaert I, De Maeseneire S, Develter D, Soetaert W, Vandamme E. Cloning and characterisation of the glyceraldehyde 3-phosphate dehydrogenase gene of Candida bombicola and use of its promoter. JOURNAL OF INDUSTRIAL MICROBIOLOGY AND BIOTECHNOLOGY. 2008;35(10):1085–92.
MLA
Van Bogaert, Inge, Sofie De Maeseneire, Dirk Develter, et al. “Cloning and Characterisation of the Glyceraldehyde 3-phosphate Dehydrogenase Gene of Candida Bombicola and Use of Its Promoter.” JOURNAL OF INDUSTRIAL MICROBIOLOGY AND BIOTECHNOLOGY 35.10 (2008): 1085–1092. Print.