title: Expression and promoter characterization of BbPacC, a pH response transcription factor gene of the entomopathogenic fungus Beauveria bassiana.
abstract: To survive, the entomopathogenic fungus Beauveria bassiana, which shows promise as a biocontrol agent for a variety of pests, including agricultural and forestry pests and vectors of human pathogens, must tailor gene expression to the particular pH of its environment. The pH response transcription factor gene BbPacC and its flanking sequence were cloned from this fungus. Quantitative reverse transcription (RT)-PCR revealed that it is highly induced by alkaline pH and salt stress, and the expression level achieved twice that of the housekeeping gene γ-actin. A microfluorometric assay indicated that the 1479 bp promoter region could activate the expression of enhanced green fluorescent protein (EGFP) under the same conditions. Truncation analysis showed that the 1479, 1274, 1040, 888 and 742 bp promoters have similar efficiencies in activating expression of β-glucuronidase (GUS). The GUS activities of corresponding transformants reached approximately 50 % that of those containing the strong constitutive promoter PtrpC. A truncation upstream at the -572 bp position (referenced to the translation start codon ATG), however, resulted in a significant loss of GUS activity. Both the upstream absences of the -502 and -387 bp positions caused almost complete loss of GUS activity. These results suggest that PPacC is an efficient, alkaline, and salt-inducible promoter, the core cis-elements are mainly located within the -742 to -502 bp region, and promoters equal to or longer than 742 bp may be feasible for regulating gene expression in response to an ambient pH or salt stress.
authors: Biotechnology Research Center, Southwest University, 215 Tiansheng Road, Beibei 400716, Chongqing, PR China.
authors: Yong-Hong
authors: YH
authors: Zhou
authors: Biotechnology Research Center, Southwest University, 215 Tiansheng Road, Beibei 400716, Chongqing, PR China.
authors: Lei
authors: L
authors: Hou
authors: Biotechnology Research Center, Southwest University, 215 Tiansheng Road, Beibei 400716, Chongqing, PR China.
authors: Yong-Jun
authors: YJ
authors: Zhang
authors: Biotechnology Research Center, Southwest University, 215 Tiansheng Road, Beibei 400716, Chongqing, PR China.
authors: Yan-Hua
authors: YH
authors: Fan
authors: Biotechnology Research Center, Southwest University, 215 Tiansheng Road, Beibei 400716, Chongqing, PR China.
authors: Zhi-Bing
authors: ZB
authors: Luo
authors: Biotechnology Research Center, Southwest University, 215 Tiansheng Road, Beibei 400716, Chongqing, PR China.
authors: Dan
authors: D
authors: Jin
authors: Biotechnology Research Center, Southwest University, 215 Tiansheng Road, Beibei 400716, Chongqing, PR China.
authors: Qiao-Sheng
authors: QS
authors: Zhou
authors: Biotechnology Research Center, Southwest University, 215 Tiansheng Road, Beibei 400716, Chongqing, PR China.
authors: Yu-Jie
authors: YJ
authors: Li
authors: Biotechnology Research Center, Southwest University, 215 Tiansheng Road, Beibei 400716, Chongqing, PR China.
authors: You
authors: Y
authors: Wang
authors: Biotechnology Research Center, Southwest University, 215 Tiansheng Road, Beibei 400716, Chongqing, PR China.
authors: Yan
authors: Y
authors: Pei
mesh_terms: D052982:Beauveria / Q000187:drug effects* / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D004252:DNA Mutational Analysis; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D020869:Gene Expression Profiling; D015966:Gene Expression Regulation, Fungal; D017930:Genes, Reporter; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D060888:Real-Time Polymerase Chain Reaction; D012492:Salts; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion; D014157:Transcription Factors / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D004271:DNA, Fungal; D012492:Salts; D014157:Transcription Factors