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Title: Process for selection of Oxygen-tolerant algal mutants that produce H.sub.2

Abstract

A process for selection of oxygen-tolerant, H.sub.2 -producing algal mutant cells comprising: (a) growing algal cells photoautotrophically under fluorescent light to mid log phase; (b) inducing algal cells grown photoautrophically under fluorescent light to mid log phase in step (a) anaerobically by (1) resuspending the cells in a buffer solution and making said suspension anaerobic with an inert gas; (2) incubating the suspension in the absence of light at ambient temperature; (c) treating the cells from step (b) with metronidazole, sodium azide, and added oxygen to controlled concentrations in the presence of white light. (d) washing off metronidazole and sodium azide to obtain final cell suspension; (e) plating said final cell suspension on a minimal medium and incubating in light at a temperature sufficient to enable colonies to appear; (f) counting the number of colonies to determine the percent of mutant survivors; and (g) testing survivors to identify oxygen-tolerant H.sub.2 -producing mutants.

Inventors:
 [1];  [1]
  1. Lakewood, CO
Issue Date:
Research Org.:
Midwest Research Institute, Kansas City, MO (United States)
OSTI Identifier:
872155
Patent Number(s):
5871952
Assignee:
Midwest Research Institute (Kansas City, MO)
Patent Classifications (CPCs):
C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES
C - CHEMISTRY C12 - BIOCHEMISTRY C12Q - MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
DOE Contract Number:  
AC36-83CH10093
Resource Type:
Patent
Country of Publication:
United States
Language:
English
Subject:
process; selection; oxygen-tolerant; algal; mutants; produce; -producing; mutant; cells; comprising; growing; photoautotrophically; fluorescent; light; mid; log; phase; inducing; grown; photoautrophically; step; anaerobically; resuspending; buffer; solution; suspension; anaerobic; inert; gas; incubating; absence; ambient; temperature; treating; metronidazole; sodium; azide; added; oxygen; controlled; concentrations; presence; white; washing; obtain; final; cell; plating; minimal; medium; sufficient; enable; colonies; appear; counting; determine; percent; survivors; testing; identify; fluorescent light; inert gas; ambient temperature; temperature sufficient; white light; buffer solution; mutant cells; algal mutant; cells grown; cells grow; cells comprising; /435/

Citation Formats

Ghirardi, Maria L, and Seibert, Michael. Process for selection of Oxygen-tolerant algal mutants that produce H.sub.2. United States: N. p., 1999. Web.
Ghirardi, Maria L, & Seibert, Michael. Process for selection of Oxygen-tolerant algal mutants that produce H.sub.2. United States.
Ghirardi, Maria L, and Seibert, Michael. Fri . "Process for selection of Oxygen-tolerant algal mutants that produce H.sub.2". United States. https://www.osti.gov/servlets/purl/872155.
@article{osti_872155,
title = {Process for selection of Oxygen-tolerant algal mutants that produce H.sub.2},
author = {Ghirardi, Maria L and Seibert, Michael},
abstractNote = {A process for selection of oxygen-tolerant, H.sub.2 -producing algal mutant cells comprising: (a) growing algal cells photoautotrophically under fluorescent light to mid log phase; (b) inducing algal cells grown photoautrophically under fluorescent light to mid log phase in step (a) anaerobically by (1) resuspending the cells in a buffer solution and making said suspension anaerobic with an inert gas; (2) incubating the suspension in the absence of light at ambient temperature; (c) treating the cells from step (b) with metronidazole, sodium azide, and added oxygen to controlled concentrations in the presence of white light. (d) washing off metronidazole and sodium azide to obtain final cell suspension; (e) plating said final cell suspension on a minimal medium and incubating in light at a temperature sufficient to enable colonies to appear; (f) counting the number of colonies to determine the percent of mutant survivors; and (g) testing survivors to identify oxygen-tolerant H.sub.2 -producing mutants.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {Fri Jan 01 00:00:00 EST 1999},
month = {Fri Jan 01 00:00:00 EST 1999}
}

Works referenced in this record:

Activation and De Novo Synthesis of Hydrogenase in Chlamydomonas
journal, December 1984


Induction, localization and metal content of hydrogenase in the green alga Chlamydomonas reinhardtii
journal, June 1994


High-frequency nuclear transformation of Chlamydomonas reinhardtii.
journal, February 1990


A rapid procedure for selective enrichment of photosynthetic electron transport mutants.
journal, February 1977


Anionic modulation of the catalytic activity of hydrogenase from Chlamydomonas reinhardtii
journal, January 1982