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首页> 外文期刊>African Journal of Biotechnology >Cloning and expression of a small heat shock protein gene CaHSP24 from pepper under abiotic stress
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Cloning and expression of a small heat shock protein gene CaHSP24 from pepper under abiotic stress

机译：非生物胁迫下辣椒小热激蛋白基因CaHSP24的克隆与表达

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摘要

The sequence of a small heat shock protein (sHSP) gene?CaHSP24?was obtained byhomology-based candidate gene method and rapid amplification of cDNA ends (RACE). The cDNA sequence of this gene is 920 bp in size (GenBank: HM132040) and contains an open reading frame (ORF) of 636 bp, which was predicted to encode a protein with 211 amino acid residues. The phylogenetic tree showed that?CaHSP24was quite similar to mitochondrial sHSPs from other plants but was distantly related to sHSPs of pepper cytoplasm and chloroplast. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) showed that?CaHSP24?was hardly detectable at 32°C, but accumulated markedly at 40°C. The gene was expressed 0.5 h after exposure to heat stress and the expression reached a peak in 1.5 h; the expression level in heat-resistant cultivar B35 was higher than that in heat-sensitive cultivar B6. The gene was also expressed weakly under salinity, heavy metal, low temperature and oxidative stresses; the expression levels under these conditions were remarkably lower than those under heat stress. Cell viability experiments showed that the heterologous expression of?CaHSP24?could enhance the viability ofEscherichia coli?under heat stress. To sum up,?CaHSP24?may play an important role for response to heat stress condition.
机译：通过基于同源性的候选基因方法并快速扩增cDNA末端（RACE），获得了一个小的热休克蛋白（sHSP）基因？CaHSP24？的序列。该基因的cDNA序列大小为920 bp（GenBank：HM132040），并包含一个636 bp的开放阅读框（ORF），据预测该编码框可编码具有211个氨基酸残基的蛋白质。系统进化树表明，？CaHSP24与其他植物的线粒体sHSP非常相似，但与辣椒细胞质和叶绿体的sHSP有很远的联系。半定量逆转录聚合酶链反应（RT-PCR）表明，CaHSP24α在32℃时几乎检测不到，但在40℃时显着积累。暴露于热应激后0.5小时，该基因表达，并在1.5小时内达到峰值。耐热品种B35的表达水平高于热敏品种B6的表达水平。该基因在盐度，重金属，低温和氧化胁迫下也表达较弱。这些条件下的表达水平明显低于热应激条件下的表达水平。细胞活力实验表明，CaHSP24β的异源表达可以增强大肠埃希菌在热胁迫下的生存能力。综上所述，CaHSP24可能在热应激条件下起重要作用。

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