Characterization of a highly active promoter, PBbgpd, in Beauveria bassiana.
The promoter of glyceraldehyde-3-phosphate dehydrogenase (gpd) gene from Aspergillus nidulans (PgpdA) is widely used to direct expression of target genes constitutively in fungi. However, in some species, a heterogeneous promoter is found to be of low efficiency. To obtain a high-efficiency promoter for transformation of Beauveria bassiana, an entomopathogenic fungus widely used as an mycoinsecticide, a glyceraldehyde-3-phosphate dehydrogenase gene (Bbgpd) promoter, was cloned and characterized. Four deletion constructs (-2118, -1153, -726, and -354) of the 5'-upstream sequence of Bbgpd linked to a bar::gus fusion gene (phosphinothricin-resistance::beta-glucuronidase fused gene), which were used as selected marker gene and report gene, respectively, were generated. GUS activities of transgenic strains harboring -726, -1153, and -2118 deletion constructs were much stronger than that of the promoter of Aspergillus nidulans gpdA (PgpdA), with a twofold to threefold increase over that in the PgpdA construct. The -726 fragment was necessary to direct GUS expression in B. bassiana. No -354 transgenic progenies were obtained, possibly because it failed to initiate the transcription of bar::gus fusion gene. A remarkable increase of GUS activity was found between the -1153 and -726 constructs, indicating that some active transcriptional elements were located in this region. With a high expression level and relatively short sequence, PBbgpd can be used to drive target genes in B. bassiana transgenic research.
Liao
Xing-gang
XG
Biotechnology Research Center, Key Laboratory of Biotechnology & Crop Quality Improvement, Ministry of Agriculture, Southwest University, 215 Tiansheng Road, Beibei, Chongqing 400715, People's Republic of China.
Fang
Wei-guo
WG
Zhang
Yong-jun
YJ
Fan
Yan-hua
YH
Wu
Xing-wei
XW
Zhou
Qun
Q
Pei
Yan
Y
D024506:5' Flanking Region; D000595:Amino Acid Sequence; D001483:Base Sequence; D052982:Beauveria / Q000235:genetics* / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics; D015870:Gene Expression; D017930:Genes, Reporter; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D017384:Sequence Deletion
D005656:Fungal Proteins; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D005966:Glucuronidase