beta-D-glucuronidase gene from Escherichia coli is a functional reporter in the methylotrophic yeast Pichia pastoris.
The Escherichia coli gene gusA was expressed in the methylotrophic yeast Pichia pastoris in a transcriptional fusion to the homologous methanol-inducible AOX1 promoter. Four recombinant clones were selected for expression studies in shake flask conditions and beta-D-glucuronidase (beta-GUS) activity was assayed each 24 h during the induction period. Regardless of the genomic integration patterns and the gene dosage, beta-GUS was functionally expressed and easily detected in all studied clones. The results obtained demonstrate the feasibility of using this bacterial enzyme as a reporter in Pichia pastoris.
Ayra-Pardo
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Plant Division, Center for Genetic Engineering and Biotechnology (CIGB), Havana, Cuba. camilo.ayra@cigb.edu.cu
Montejo-Sierra
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Vázquez-Padrón
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García-Martínez
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C
D015139:Blotting, Southern; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D015870:Gene Expression; D017930:Genes, Reporter; D005966:Glucuronidase / Q000235:genetics* / Q000378:metabolism*; D024101:Mitochondrial Proteins; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism; D010641:Phenotype; D010843:Pichia / Q000201:enzymology / Q000235:genetics*; D010940:Plant Proteins; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D011995:Recombination, Genetic; D014170:Transformation, Genetic; D000099188:Alternative Oxidase
D005966:Glucuronidase; D024101:Mitochondrial Proteins; D010088:Oxidoreductases; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; D000099188:Alternative Oxidase