Sequence and expression of the gene for N10‐formyltetrahydrofolate synthetase from Clostridium cylindrosporum

Sau3 A and Hind III restriction fragments of Clostridium cylindrosporum genomic DNA were used to isolate clones containing 80% of the N10‐H4folate synthetase gene in a 5′ fragment and the remaining 20% of the gene in the 3′ fragment. These fragments were joined at a common SnaB I restriction site an...

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Bibliographic Details
Published in:Protein science 1993-02, Vol.2 (2), p.197-205
Main Authors: Rankin, Carolyn A., Haslam, Gale C., Himes, Richard H.
Format: Article
Language:English
Subjects:
Amino Acid Sequence
ATP consensus sequence
Base Sequence
Cloning, Molecular
Clostridia
Clostridium - enzymology
Clostridium - genetics
Enzyme Stability - genetics
Escherichia coli - genetics
Formate-Tetrahydrofolate Ligase - biosynthesis
Formate-Tetrahydrofolate Ligase - genetics
formyltetrahydrofolate synthetase
Gene Library
gene sequence
Genes, Bacterial - genetics
Genetic Code
Molecular Sequence Data
Protein Structure, Secondary
Sequence Analysis
Sequence Homology, Amino Acid
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Summary:Sau3 A and Hind III restriction fragments of Clostridium cylindrosporum genomic DNA were used to isolate clones containing 80% of the N10‐H4folate synthetase gene in a 5′ fragment and the remaining 20% of the gene in the 3′ fragment. These fragments were joined at a common SnaB I restriction site and expressed in Escherichia coli at a level equivalent to what is normally found in C. cylindrosporum. Sequence comparisons show a large degree of homology with genes from two other clostridial species, including a thermophile. Certain conserved sequences found in the three clostridial proteins and in the N10‐H4folate synthetase portion of eukaryotic Cl‐H4folate synthases may represent consensus sequences for nucleotide and H4folate binding.
ISSN:0961-8368
1469-896X
DOI:10.1002/pro.5560020208