Rapid transformation of nor-Saccharomyces yeasts by electroporation

Martínez, Eduardo Justo; García, Carmen Gallo; Grillo, Juan Morales

doi:10.1007/bf00156369

Cited by 6 publications

(3 citation statements)

References 6 publications

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“…Transformation of Pichia pastoris and screening for Mut S phenotype The expression plasmid was linearized with ClaI and it was used to transform P. pastoris strain MP36 (HIS3) (Yong et al 1992) by electroporation (Martínez et al 1993). The transformation mixture was plated on selective medium [1.34 % (w/v) yeast nitrogen base, 3 % (w/v) agar, 1 M sorbitol, 2 % (w/v) glucose].…”

Section: Construction Of the Expression Vector Pp-tighmentioning

confidence: 99%

Pichia pastoris expressing recombinant tilapia growth hormone accelerates the growth of tilapia

Acosta

Morales

et al. 2007

Biotechnol Lett

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Section: Construction Of the Expression Vector Pp-tighmentioning

confidence: 99%

Pichia pastoris expressing recombinant tilapia growth hormone accelerates the growth of tilapia

Acosta

Morales

et al. 2007

Biotechnol Lett

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“…P. pastoris cells were transformed by the colony‐electroporation procedure previously described by Martinez et al . (1993) .…”

Section: Methodsmentioning

confidence: 99%

“…P. pastoris cells were transformed by the colony-electroporation procedure previously described by Martinez et al (1993). The resulting transformants (PPGUS) were selected on YPD plates supplemented with 200 mg ml −1 of G418 and immediately replicated onto YPD plates containing 1 mg ml −1 of G418 for 'Jackpot' clones screening.…”

Section: Transformation and Selectionmentioning

confidence: 99%

bb-D-glucuronidase gene from Escherichia coli is a functional reporter in the methylotrophic yeast Pichia pastoris

Ayra‐Pardo

Montejo‐Sierra

ázquez‐Padrón

et al. 1999

Lett Appl Microbiol

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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 β‐ d‐glucuronidase (β‐GUS) activity was assayed each 24 h during the induction period. Regardless of the genomic integration patterns and the gene dosage, β‐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.

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Multimeric HCV E2 protein obtained from pichia pastoris cells induces a strong immune response in mice

Martínez‐Donato

Capdesuñer²,

Acosta‐Rivero³

et al. 2007

Mol Biotechnol

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Production of immunogenic hepatitis C virus (HCV) envelope proteins will assist in the future development of preventive or therapeutics applications. Only properly folded monomeric E2 protein is able to bind a putative cellular co-receptor CD81, but this interaction may modulate cell immune function. Recombinant E2 proteins, similar to the native form, but lacking undesirable immunoregulatory features, might be promising components of vaccine candidates against HCV. To obtain E2 suitable for structural as well as functional studies, a recombinant E2 variant (E2680) was produced in Pichia pastoris cells. E2680, comprising amino acids 384 to 680 of the HCV polyprotein, was secreted into the culture supernatant in the N-glycosilated form and was mainly composed of disulfide-linked multimers. Both monomeric and oligomeric forms of E2680 were recognized by conformational-sensitive MAb H53. In addition, antibodies in sera from 70% of HCVpositive patients were reactive against E2680. By immunizing E2680 in BALB/c mice, both a specific cellular immune response and anti-E2680 IgG antibody titers of 1:200,000 were induced. Our data suggest that recombinant E2680 could be useful to successfully induce strong anti-HCV immunity.

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Rapid transformation of nor-Saccharomyces yeasts by electroporation

Cited by 6 publications

References 6 publications

Pichia pastoris expressing recombinant tilapia growth hormone accelerates the growth of tilapia

Pichia pastoris expressing recombinant tilapia growth hormone accelerates the growth of tilapia

bb-D-glucuronidase gene from Escherichia coli is a functional reporter in the methylotrophic yeast Pichia pastoris

Multimeric HCV E2 protein obtained from pichia pastoris cells induces a strong immune response in mice

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