Genetics of the methanogenic bacterium, Methanococcus voltae, with attention to genetic control mechanisms: Progress report, January 15, 1988--January 14, 1989

Technical Report · Wed Feb 01 04:00:00 EST 1989

OSTI ID:6800239

Konisky, J

The overall objective of our program is to both develop a genetic system and understand mechanisms of gene expression for the methanogen, Methanococcus voltae. During the past year we have made progress in several areas. During the last several months, we have had very promising results with regard to the utilization of electroporation for the introduction of DNA into Mc.voltae. In summary, we have been able to demonstrate an approximately 2 to 5 fold increase in transformation via electroporation as compared to the frequency obtained via natural transformation. In the coming year, we intend to further optimize this system and to make a serious effort to introduce plasmid and VLP genome DNA. In our efforts to characterize gene expression in methanogens, we have collaborated with T. Demarco and Ralph Wolfe in cloning an enzyme of the C1 cycle. To catalyze the interconversions mediated by the novel coenzymes of methanogenesis, several enzymes exist which have no common analog in non-methanogenic bacteria. One of these enzymes, formylmethanofuran: tetahydromethanopter in fiormyltransferase was cloned into E. coli HB101. Methanobacterium thermoautotrophicum (H) DNA digested with BAM H1 was ligated into pBR329 and probed with a synthetic oligonucleotide directed against the portion of the gene encoding the amino terminus of the enzyme. The cloned 5.0-kilobase genomic insert contained the entire gene for this polypeptide as well as 200 bp of the upstream region. A ribosome binding site just upstream of the initiation codon was identified, however a promoter was not clearly visible owing to the naturally high A plus T content of this intergenic region. 4 refs.

• Research Organization: Illinois Univ., Urbana (USA). Dept. of Microbiology
• DOE Contract Number: FG02-84ER13241
• OSTI ID: 6800239
• Report Number(s): DOE/ER/13241-1; ON: DE89006249
• Country of Publication: United States
• Language: English

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59 BASIC BIOLOGICAL SCIENCES
ALKANES
BACTERIA
BIOLOGY
CELL CONSTITUENTS
CLONING
DNA HYBRIDIZATION
DNA-CLONING
DOCUMENT TYPES
ENZYME ACTIVITY
GENE OPERONS
GENE REGULATION
GENETICS
GROWTH
HYBRIDIZATION
HYDROCARBONS
METHANE
METHANOGENIC BACTERIA
MICROORGANISMS
ORGANIC COMPOUNDS
PLASMIDS
PRODUCTION
PROGRESS REPORT