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Na+/K+-ATPase Alpha Subunit in the Monogonont Rotifer, Brachionus koreanus: Molecular Cloning and Response to Different Salinity KCI 등재
- 언어ENG
- URLhttps://db.koreascholar.com/Article/Detail/315820
Na+/K+-ATPase is a membrane protein and plays a key role in osmotic regulation in living organisms. In the present study, a cDNA sequence encoding the Na+/K+-ATPase alpha subunit from the monogonont rotifer, Brachionus koreanus was cloned by rapid amplification of cDNA ends technique. To investigate the role of this enzyme in osmotic stress, enzymatic activities of Na+/K+-ATPase were measured after exposure to different salinities for 48 h. The full-length Bk Na+/K+-ATPase cDNA was 3069 bp-long, encoding a 1022-amino acid polypeptide. Bk Na+/K+- ATPase possesses eight membrane spanning regions and five conserved domains. Phylogenetic analysis showed that Bk Na+/K+-ATPase had high identity with those of other species, and was closely clustered with other Brachionus sp. These findings indicate that this protein was conserved both structurally and functionally. B. koreanus Na+/K+-ATPase activity was stimulated in both hyposaline (6 psu) and hypersaline (32 psu) conditions, suggesting that this protein may play a role in osmoregulation. This study would provide better understanding of the physiology of B. koreanus and this enzyme may be useful as a molecular marker for evaluation of osmotic stress in aquatic environment.
Materials and methods
1. Culture and maintenance
2. Cloning and sequence analysis
3. Phylogenetic analysis
4. Measurement of Na+/K+-ATPase activity indifferent salinity
5. Statistical analysis
Results
1. C loning and sequence analysis of the B. koreanusNa+/K+-ATPase alpha subunit
2. Phylogenetic relationships
3. Altered Na+/K+-ATPase activity in response tosalinity
Discussion
References
