Plasmids for expression of heterologous proteins in Rhizopus oryzae.

Plasmids for expression of heterologous proteins in Rhizopus oryzae. Research Abstract Details 

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Plasmids for expression of heterologous proteins in Rhizopus oryzae. Abstract Text:

Jeffrey A Mertens,Christopher D Skory,Ashraf S Ibrahim,

Rhizopus oryzae has long been used for enzyme production (e.g., glucoamylase and lipase), organic acid synthesis, and various fermented food applications. In this work, we describe a set of plasmid-based expression vectors that can be used for the production of heterologous proteins in R. oryzae. Three plasmid vectors have been created using either the glucoamylase A (amyA), pyruvate decarboxylase (pdcA), or phosphoglycerate kinase (pgk1) promoters to drive expression of heterologous proteins. All three plasmids use the pdcA terminator for transcription termination, the pyrG gene for restoration of uracil prototrophy, and an ampicillin resistance gene and origin of replication for maintenance in Escherichia coli. We have expressed green fluorescent protein (GFP) and compared transcription and protein accumulation for each of the expression vectors. Accumulation of GFP transcript and protein was directly correlated with the choice of promoter with pdcA > amyA > pgk1. Transcript level appears to parallel GFP protein accumulation. Plasmid copy number had little impact on transcription or protein accumulation. These vectors should be useful for overexpression of heterologous proteins and potentially, metabolic engineering of Rhizopus strains.

Plasmids for expression of heterologous proteins in Rhizopus oryzae. Publishing Authors By Initials

JA Mertens,CD Skory,AS Ibrahim,

For similar genetic processes: recombination, genetic: transformation, genetic research abstracts see: genetic processes: recombination, genetic: transformation, genetic research

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Plasmids for expression of heterologous proteins in Rhizopus oryzae. Journal Published:

PUBLICATION TYPE: Research Support, N.I.H., Extr

Journal: Archives of microbiology

VOLUME: 186

Page Numbers: 41-50

Journal Abbreviation: Arch. Microbiol.

ISSN: 0302-8933

DAY: 28

MONTH: 06

YEAR: 2006

Plasmids for expression of heterologous proteins in Rhizopus oryzae. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 410427

Plasmids for expression of heterologous proteins in Rhizopus oryzae. Keywords Mesh Terms:

KEYWORDS: Transformation, Genetic

MESH TERMS: metabolism

Chemical & Substance for Abstract: Plasmids for expression of heterologous proteins in Rhizopus oryzae. Information

Substance Name: Green Fluorescent Proteins

Registry Number: 147336-22-9

Grant and Affiliation Information for Plasmids for expression of heterologous proteins in Rhizopus oryzae.

AFFILIATION: Fermentation Biotechnology Research Unit, National Center for Agricultural Utilization Research, USDA, Agricultural Research Service, 1815 N. University St., Peoria, IL 61604, USA. mertensj@ncaur.usda.gov

Country: Germany

AGENCY: United States NIAID

GRANT: R56 AI063503-01A1

ACRONYM: AI

MEDLINETA: Arch Microbiol

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