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Isolation and Characterization of a Rice Mitochondrial Small Heat Shock Protein Gene KCI 등재
- 언어ENG
- URLhttps://db.koreascholar.com/Article/Detail/403849
pp.285-290
To understand the role of small heat shock protein (sHSPs) in rice plant response to various stresses such as the heat and oxidative stresses, a cDNA encoding a 24.1 kDa mitochondrial small HSP (Oshsp24.1) was isolated from rice by rapid amplification of cDNA ends (RACE) PCR. The deduced amino acid sequence shows very high similarity with other plant small HSPs. DNA gel blot analysis suggests that the rice genome contains more than one copy of Oshsp24.1. High level of expression of Oshsp24.1 transcript was observed in rice seedlings in response to heat, methyl viologen, hydrogen peroxide, ozone, salt and heavy metal stresses. Recombinant OsHSP24.1 protein was produced in E. coli cells for biochemical assay. The protein formed oligomeric complex when incubated with Sulfo-EGS (ethylene glycol bis (succinimidyl succinate)). Our results shows that Oshsp24.1 has an important role in abiotic stress response and have potential for developing stress-tolerant plants.
Ⅰ. INTRODUCTION
Ⅱ. MATERIALS AND METHODS
1. Plant growth and stress treatment
2. DNA and RNA gel blot analysis
3. Cloning of Oshsp24.1 full-length cDNA by RACE-PCR
4. Expression of OsHSP24.1 recombinant protein in E.coli
5. Cross-linking and immunobloting
Ⅲ. Results and Discussion
1. Cloning of a Oshsp24.1 cDNA from rice
2. Characterization of the Oshsp24.1 sequence
3. Genomic organization of the Oshsp24.1 gene
4. Expression of Oshsp24.1 in response to abiotic stresses
5. Biochemical characterization of recombinant OsHSP24.1protein
Ⅳ. Acknowledgement
Ⅴ. REFERENCES
