Expression of HSP70 in response to heat-shock and its cDNA cloning from Mediterranean blue mussel
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Expression of HSP70 in response to heat-shock was investigated at the protein and mRNA levels in Mediterranean blue mussel. Western and Northern blot analyses revealed that HSP70 was expressed following heat-shock in the mantle at both protein and mRNA levels, suggesting that gene expression of HSP70 is implicated in the cellular response to heat-shock stress in mussel. It was then attempted to clone HSP70 cDNA in order to determine the primary structure of mussel HSP70. As a result, two full-length cDNA encoding HSP70 were isolated from a cDNA library prepared from the heat-shocked mantle. The isolated cDNA consist of single open reading frames of 2067 bp and 1911 bp which encode proteins of 689 amino acids and 637 amino acids, respectively. Both HSP70 cDNA encode an ATPase do main, and a substrate-binding do main in addition to a Glu-Glu-Val-Asp (EEVD) peptide motif that is specific for cytosolic HSP70. These findings suggest that the cDNA clones obtained in the present study encode cytosolic HSP70.
Key Wordsheat-shock HSP70 mantle Mediterranean blue mussel Mytilus galloprovin-cialis stress
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- 2.Morimoto RI, Tissieres A, Georgopoulos C. Stress Protein in Biology and Medicine, Cold Spring Harbour Laboratory Press, New York, 1994.Google Scholar
- 4.Somero GN, Hofman GE. Temperature thresholds for protein adaptation: when does temperaturt change start to ‘hurt’? In: Wood CM, McDonald DG (eds). Global Warming: Implications for Freshwater and Marine Fish. Cambridge University Press, Cambridge, 1996; 1–24Google Scholar
- 8.Sanders BM, Pascoe VM, Nakagawa PA, Martin LS. Persistence of the heat-shock response over time in a common Mytilus mussel. Mol. Mar. Biol. Biotech. 1992; 1: 147–154.Google Scholar
- 9.Hofmann GE, Somero GN. Evidence for protein damage at environmental temperature. Seasonal changes levels of ubiquitin conjugates and HSP70 in the intertidal mussel Mytilus trossulus. J. Exp. Zool. 1995; 198: 1509–1518.Google Scholar
- 11.Hofmann GE, Somero GN. Protein ubiquitination and stress protein synthesis in Mytilus trossulus occurs during recovery from tidal emersion. Mol. Mar. Biol. Biotech. 1996; 5: 175–184.Google Scholar
- 20.Sambrook J, Russel DW. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, New York, 2001.Google Scholar