Cloning, characterization and application of a glyceraldehyde-3-phosphate dehydrogenase promoter from Aspergillusterreus

Short Communication

DOI: 10.1007/s10295-013-1385-0

Cite this article as:
Huang, X., Lu, X. & Li, JJ. J Ind Microbiol Biotechnol (2014) 41: 585. doi:10.1007/s10295-013-1385-0

Abstract

It is important to develop native and highly efficient promoters for effective genetic engineering of filamentous fungi. Although Aspergillusterreus is an important industrial fungus for the production of itaconic acid and lovastatin, the available genetic toolbox for this microorganism is still rather limited. We have cloned the 5′ upstream region of the glyceraldehyde-3-phosphate dehydrogenase gene (gpd; 2,150 bp from the start codon) from A. terreus CICC 40205 and subsequently confirmed its promoter function using sgfp (synthetic green fluorescent protein) as the reporter. The sequence of the promoter PgpdAt was further analysed by systematic deletion to obtain an effective and compact functional promoter. Two truncated versions of PgpdAt (1,081 and 630 bp) were also able to drive sgfp expression in A. terreus. The activities of these three PgpdAt promoters of varying different lengths were further confirmed by fluorescence, western blot and transcription. The shortest one (630 bp) was successfully applied as a driver of vgb expression in the genetic engineering of A. terreus. The function of expressed haemoglobin was demonstrated by the CO (carbon monoxide)-difference spectrum and enhanced oxygen uptake rate, glucose consumption and itaconic acid titer. Our study was successful in developing and validating an efficient and compact native promoter for genetic engineering of A. terreus.

Keywords

Aspergillusterreus Glyceraldehyde-3-phosphate dehydrogenase promoter Native promoter Promoter function Vitreoscilla haemoglobin 

Supplementary material

10295_2013_1385_MOESM1_ESM.docx (2.4 mb)
Supplementary material 1 (DOCX 2436 kb)

Copyright information

© Society for Industrial Microbiology and Biotechnology 2013

Authors and Affiliations

  1. 1.Key Laboratory of Biofuels, Shandong Provincial Key Laboratory of Energy Genetics, Qingdao Institute of Bioenergy and Bioprocess TechnologyChinese Academy of SciencesQingdaoChina

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