title: Key regulatory elements of a strong constitutive promoter, P GCW14, from Pichia pastoris.
abstract: The promoter of the Pichia pastoris gene GCW14 is strong and constitutive when glycerol is the available carbon source. To identify the cis-acting elements of this promoter (P GCW14), we constructed expression plasmids where the enhanced green fluorescent protein gene was fused to a series of mutants of P GCW14. We identified one negative (-114 to -94) and three positive regulatory regions (-426 to -152, -134 to -114, -94 to -77). The TATA box of P GCW14 was located at -48. One negative and four positive regulatory sites were identified combining error-prone PCR and directed mutation. The mutated promoter, M+20, with an increased promoter activity, was then used to express the gene for lipase B from Candida antarctica.
authors: School of Bioscience and Bioengineering, South China University of Technology, Guangzhou, 510006, Guangdong, China, zhangxuanwei88@yahoo.com.
authors: Xuanwei
authors: X
authors: Zhang
authors: Xiuqin
authors: X
authors: Zhang
authors: Shuli
authors: S
authors: Liang
authors: Yanrui
authors: Y
authors: Ye
authors: Ying
authors: Y
authors: Lin
mesh_terms: D005656:Fungal Proteins / Q000235:genetics; D015966:Gene Expression Regulation, Fungal / Q000235:genetics*; D005800:Genes, Fungal / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics; D008049:Lipase / Q000235:genetics / Q000378:metabolism; D051839:NFI Transcription Factors / Q000235:genetics; D010843:Pichia / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D016385:TATA Box / Q000235:genetics
chemical_list: D005656:Fungal Proteins; D051839:NFI Transcription Factors; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D008049:Lipase