Molecular Cloning and Characterization of theCaenorhabditis elegansElongation Factor 2 Gene (eft-2)
| Autor: | E. P. M. Candido, E. N. Ofulue |
|---|---|
| Rok vydání: | 1991 |
| Předmět: |
Untranslated region
Sequence analysis Molecular Sequence Data Restriction Mapping Sequence alignment Polymerase Chain Reaction Conserved sequence Peptide Elongation Factor 2 Sequence Homology Nucleic Acid Genetics Animals Amino Acid Sequence RNA Messenger Cloning Molecular Molecular Biology Peptide sequence Caenorhabditis elegans Base Sequence biology cDNA library Nucleic acid sequence DNA Cell Biology General Medicine Blotting Northern Peptide Elongation Factors biology.organism_classification Molecular biology Blotting Southern Protein Biosynthesis Caenorhabditis Sequence Alignment |
| Zdroj: | DNA and Cell Biology. 10:603-611 |
| ISSN: | 1557-7430 1044-5498 |
| DOI: | 10.1089/dna.1991.10.603 |
| Popis: | A Caenorhabditis elegans lambda ZAP cDNA library was screened using a fragment amplified from highly conserved regions of the mammalian and Drosophila elongation factor 2 (EF-2). Two types of cDNA clones were obtained, corresponding to two mRNA species with 3'-untranslated regions of 60 and 115 nucleotides, both encoding identical polypeptides. Sequence analysis of these clones and comparisons with hamster and Drosophila EF-2 sequences suggests that they encode C. elegans EF-2. Clone pCef6A, encoding the entire C. elegans EF-2 mRNA sequence including 45 nucleotides of 5'-untranslated region, contains a 2,556-bp open reading frame which predicts a polypeptide of 852 amino acid residues (Mr 94,564). The deduced amino acid sequence is greater than 80% identical to that of mammalian and Drosophila EF-2. Conserved sequence segments shared among a variety of GTP-binding proteins are found in the amino-terminal region. The carboxy-terminal half contains segments unique to EF-2 and its prokaryotic homolog, EF-G, as well as the histidyl residue which is ADP-ribosylated by diphtheria toxin. The C. elegans protein contains a 12-amino-acid insertion between positions 90 and 100, and a 13-amino-acid deletion between positions 237 and 260, relative to hamster EF-2. Partial sequencing of a genomic clone encoding the entire C. elegans EF-2 gene (named eft-2) has so far revealed two introns of 48 and 44 bp following codons Gln-191 and Gln-250, respectively. Southern and Northern blot analyses indicate that eft-2 is a single-copy gene and encodes a 3-kb mRNA species which is present throughout nematode development. |
| Databáze: | OpenAIRE |
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