Summary
A genomic library of EcoRI*-digested cellular DNA from Methanococcus voltae was constructed in an expression vector, which allows the induction of fusion proteins of the Mc. voltae gene products with an MS2 RNA polymerase N-terminal fragment. Antibodies were raised against the subunits of the methyl CoM reductase component C complex. They were used to screen clones from the genomic library for the synthesis of polypeptides carrying antigenic determinants of the reductase subunits. Plasmids containing fragments of two of the three subunit genes were isolated. The genes were shown to be adjacent to each other by hybridization against restriction fragments of cellular DNA and Mc. voltae DNA cloned in a bacteriophage lambda replacement vector. The direction of the transcription of one of the genes was established by partial sequence determination and it was shown that the two genes have a common transcript.
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Abbreviations
- bp:
-
base pairs
- SDS:
-
sodium dodecyl sulfate
- kb:
-
kilobases
References
Balch WE, Wolfe RS (1976) New Approach to the cultivation of methanogenic bacteria: 2-mercaptoethanesulfonic acid (HS-CoM)-dependent growth of Methanobacterium ruminantium in a pressurized atmosphere. Appl Environ Microbiol 32:781–791
Balch WE, Fox GE, Margrum LJ, Woese CR, Wolfe RS (1979) Methanogens: Reevaluation of a unique biological group. Microbiol Rev 43:260–296
Benton WD, Davis RW (1977) Screening λ gt recombinant clones by hybridization to single plaques in situ. Science 196:180–182
Bolivar F, Rodriguez RL, Greene PJ, Betlach MC, Heynecker HL, Boyer HW, Crosa JH, Falkow S (1977) Construction and characterization of new cloning vehicles. II A multipurpose cloning system. Gene 2:95–113
Bollschweiler C, Klein A (1982) Polypeptide synthesis in Escherichia coli directed by cloned Methanobrevibacter arboriphilus DNA. Zbl Bakt Hyg I Abt Orig C 3:101–109
Daniels DL, Schroeder JL, Szybalski W, Sanger F, Coulson AR, Hong GF, Hill DF, Petersen GB, Blattner FR (1983) Complete annotated lambda sequence. In: Hendrix RW, Roberts JW, Stahl FW, Weisberg RA (eds) Lambda II. Cold Spring Harbor Laboratory, Cold Spring Harbor, pp 519–676
Ellefson WL, Wolfe RS (1980) Role of component C in the methyl reductase system of Methanobacterium. J Biol Chem 255:8388–8389
Hamilton PT, Reeve JN (1984) Cloning and expression of archaebacterial DNA from methanogens in Escherichia coli. In: Strohl WR, Tuovinen OH (eds) Microbial chemoautotrophy. The Ohio State University Press, Columbus, Ohio, pp 291–308
Hudson L, Hay FC (1980) Practical immunology. Blackwell Scientific Publications, Oxford
Karn J, Brenner S, Barnett L, Cesareni G (1980) Novel bacteriophage λ cloning vectors. Proc Natl Acad Sci USA 77:5172–5176
Kirkegaard LH (1973) Gradient sievorptive chromatography. A focusing system for the separation of cellular components. Biochemistry 12:3627–3632
Klein A, Schnorr M (1984) Genome complexity of methanogenic bacteria. J Bacteriol 158:628–631
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature 227:680–685
Loenen WAM, Brammar WJ (1980) A bacteriophage lambda vector for cloning large DNA fragments made with several restriction enzymes. Gene 10:249–259
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning. A laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor
Maxam AM, Gilbert W (1980) Sequencing end-labeled DNA with base specific chemical cleavages. In: Methods in Enyzmology, Vol. 65: Grossman L, Moldave K (eds) Nucleic acids, part I. Academic Press, New York, pp 499–560
Reeve JN, Trun NJ, Hamilton PT (1982) Beginning genetics with methanogens. In: Hollaender A, De Moss RD, Kaplin S, Konisky J, Savage D, Wolfe RS (eds) Genetic engineering of micro-organisms for chemicals. Plenum Press, New York, pp 233–244
Remaut E, Stanssens P, Fiers W (1981) Plasmid vectors for high efficiency expression controlled by the pL promotor of coliphage lambda. Gene 15:81–93
Southern EM (1975) Detection of specific sequences among DNA fragments. J Mol Biol 98:503–517
Thomas M, Davis RW (1975) Studies on the cleavage of bacteriophage lambda DNA with Eco RI restriction endonuclease. J Mol Biol 91:315–328
Woese CR, Magrum LJ, Fox GE (1978) Archaebacteria. J Mol Evol 11:245–252
Wolfe RS (1979) Methanogens: a surprising microbial group. Antonie van Leeuwenhoek 45:353–364
Wood AG, Redborg AH, Cue DR, Whitman WB, Konisky J (1983) Complementation of argG and hisA mutations of Escherichia coli by DNA cloned from the archaebacterium Methanococcus voltae. J Bacteriol 156:19–29
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Communicated by E. Bautz
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Konheiser, U., Pasti, G., Bollschweiler, C. et al. Physical mapping of genes coding for two subunits of methyl CoM reductase component C of Methanococcus voltae . Molec Gen Genet 198, 146–152 (1984). https://doi.org/10.1007/BF00328714
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DOI: https://doi.org/10.1007/BF00328714