Isolation and functional characterization of the ASR gene from Ipomoea pes-caprae
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摘要: 通过对厚藤（Ipomoea pes-caprae （Linn.）Sweet.）cDNA文库的筛选，获得了一个编码厚藤ASR（ABA-stress-ripening）基因的全长cDNA，命名为IpASR。研究结果显示，IpASR编码区全长648 bp，共编码215个氨基酸；蛋白质等电点为5.42，分子量为24.57 kD。通过在酵母中表达，发现IpASR能够提高转基因酵母的耐盐性及抗氧化能力。进一步以厚藤成年植株及幼苗为材料进行实时荧光定量PCR分析，结果表明，IpASR基因在厚藤成年植株各组织中广泛表达；高盐、甘露醇胁迫和ABA处理可诱导该基因在厚藤幼苗中的表达。结合GFP融合蛋白的亚细胞定位和生物信息学分析，发现IpASR蛋白为核蛋白，推测IpASR基因参与了厚藤生长发育的调控，并可响应ABA和非生物胁迫的诱导。Abstract: We focused on the full-length cDNA encoding ASR protein isolated from a Ipomoea pes-caprae (Linn.) Sweet. cDNA library. Results showed the coding region of IpASR cDNA was 648 bp, encoding a 215 amino acid protein with a molecular weight of 24.57 kD and isoelectric point of 5.42. By ectopic expression of IpASR in yeast, we found that IpASR improved the salt and H2O2 tolerance of transgenic yeast strains. Using adult plants and seedlings of I. pes-caprae with or without abiotic stress and ABA treatment, real-time RT-PCR analysis showed that IpASR was widely expressed in different adult organs in I. pes-caprae. The IpASR transcript was induced under abiotic stress and ABA treatment. The subcellular localization assay combining bioinformatics analysis showed that IpASR was a nucleoprotein. These results suggest that IpASR might play an important role in the regulation of I. pes-caprae growth and development, and therefore respond to environmental abiotic stress and the ABA signal pathway.
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Keywords:
- Ipomoea pes-caprae /
- ASR /
- Abiotic stress /
- Subcellular localization
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