Cloning and sequencing of a molluscan endo-beta-1,4-glucanase gene from the blue mussel, Mytilus edulis.
Using polymerase chain reaction, cloning and sequencing techniques, a complementary DNA encoding a low molecular mass cellulase (endo-1,4-beta-D-glucanase, EC 3.2.1.4) has been identified in the digestive gland of the marine mussel, Mytilus edulis. It contains a 5' untranslated region, a 633-nucleotide ORF encoding a 211 amino-acid protein, including a 17 amino-acid signal peptide and a complete 3' untranslated region. At the C-terminal end of the purified mature protein, a 13 amino-acid peptide is lacking in comparison to the protein sequence deduced from the ORF. This peptide is probably removed as a consequence of post-translational amidation of the C-terminal glutamine. The endoglucanase genes have been isolated and sequenced from both Swedish and French mussels. The coding parts of these two sequences are identical. Both genes contain two introns, the positions of which are conserved. However the length of the introns are different due to base substitutions, insertions or deletions showing the existence of interspecies length polymorphism. The percentage of similarity for the introns of the two gene sequences is 96.9%. This is the first time a molluscan cellulase is characterized at DNA level. Amino acid sequence-based classification has revealed that the enzyme belongs to the glycosyl hydrolase family 45 [B. Henrissat (Centre de Recherches sur les Macromolecules Végétales, CNRS, Joseph Fourier Université, Grenoble, France), personal communication]. There is no cellulose binding domain associated with the sequence.
Xu
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Center for Surface Biotechnology, Uppsala Biomedical Center, Uppsala University, Sweden.
Janson
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JC
Sellos
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D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D049872:Bivalvia / Q000201:enzymology* / Q000235:genetics*; D002480:Cellulase / Q000737:chemistry* / Q000235:genetics*; D003001:Cloning, Molecular; D017931:DNA Primers; D008969:Molecular Sequence Data; D012275:Oryza / Q000201:enzymology; D010947:Plants, Toxic; D016133:Polymerase Chain Reaction; D021382:Protein Sorting Signals; D011994:Recombinant Proteins / Q000737:chemistry; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D016385:TATA Box; D014026:Nicotiana / Q000201:enzymology; D014242:Trichoderma / Q000201:enzymology
D017931:DNA Primers; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D002480:Cellulase