Cloning and Nucleotide Sequence Determination of the Clostridium pasteurianum Ferredoxin Gene

We have constructed a library of Clostridium pasteurianum DNA cloned in the plasmid pBR322. Based on the known amino acid sequence for C. pasteurianum ferredoxin, a 64-fold degenerate heptadecanucleotide pool was synthesized. This mixed probe hybridized to two clones which were shown to contain >...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1985-03, Vol.82 (6), p.1653-1657
Main Authors: Graves, Mary C., Mullenbach, Guy T., Rabinowitz, Jesse C.
Format: Article
Language:English
Subjects:
Base Sequence
Biological and medical sciences
Biotechnology
Cloning, Molecular
Clostridium - genetics
Clostridium pasteurianum
Codon - genetics
DNA
DNA, Bacterial - genetics
Ferredoxins - genetics
Fundamental and applied biological sciences. Psychology
Gels
Genes, Bacterial
Genetic engineering
Genetic technics
Genomics
Messenger RNA
Methods. Procedures. Technologies
Molecular cloning
Nucleotide sequences
Nucleotides
Oligonucleotide probes
Open reading frames
Operon
Plasmids
Promoter regions
Protein Biosynthesis
Transcription, Genetic
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Summary:We have constructed a library of Clostridium pasteurianum DNA cloned in the plasmid pBR322. Based on the known amino acid sequence for C. pasteurianum ferredoxin, a 64-fold degenerate heptadecanucleotide pool was synthesized. This mixed probe hybridized to two clones which were shown to contain >6 kilobase pairs of the same genomic DNA. Sequence analysis of a common Sau3A1 0.6-kilobasepair fragment revealed that it contains the information for the apoferredoxin structural gene. According to the DNA sequence, the only post-translational processing of this small apoprotein is the hydrolysis of the initiator methionine. Putative transcription and translation start and stop signals are present within the sequence.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.82.6.1653