Cloning, Sequencing and Expression of a Sialidase Gene from Clostridium sordellii G12
| Autor: | Peter Roggentin, Helmut Blöcker, Roland Schauer, Bernd Rothe, Ronald Frank |
|---|---|
| Rok vydání: | 1989 |
| Předmět: |
DNA
Bacterial Molecular Sequence Data Restriction Mapping Neuraminidase Clostridium sordellii Molecular cloning Sialidase Microbiology Gene Expression Regulation Enzymologic Restriction fragment Sequence Homology Nucleic Acid Escherichia coli Consensus sequence Amino Acid Sequence Cloning Molecular Peptide sequence Clostridium Base Sequence biology Structural gene Nucleic acid sequence Gene Expression Regulation Bacterial biology.organism_classification Molecular biology Biochemistry Genes Bacterial biology.protein |
| Zdroj: | Microbiology. 135:3087-3096 |
| ISSN: | 1465-2080 1350-0872 |
| Popis: | Summary: A 4·3 kb Xbal restriction fragment of DNA from Clostridium sordellii G12 hybridized with a synthetic oligonucleotide representing the N-terminus of the sialidase protein secreted by C. sordellii. This cloned fragment was shown to encode only part of the sialidase protein. The sialidase gene of C. sordellii was completed by a 0·7 kb Rsal restriction fragment overlapping one end of the XbaI fragment. After combining the two fragments and transformation of Escherichia coli, a clone that expressed sialidase was obtained. The nucleotide sequence of the sialidase gene of C. sordellii G12 was determined. The sequence of the 18 N-terminal amino acids of the purified extracellular enzyme perfectly matched the predicted amino acid sequence near the beginning of the structural gene. The amino acid sequence derived from the complete gene corresponds to a protein with a molecular mass of 44735 Da. Upstream from the putative ATG initiation codon, ribosomal-binding site and promoter-like consensus sequences were found. The encoded protein has a leader sequence of 27 amino acids. The enzyme expressed in E. coli has similar properties to the enzyme isolated from C. sordellii, except for small differences in size and isoelectric point. Significant homology (70%) was found with a sialidase gene from C. perfringens. |
| Databáze: | OpenAIRE |
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