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OALib Journal期刊
ISSN: 2333-9721
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海马齿小热激蛋白sphsp18.1基因的克隆及盐胁迫表达分析
DOI: 10.3724/SP.J.1145.2015.04062, PP. 882-886
Keywords: 海马齿,小分子热激蛋白,分子克隆,盐胁迫
Abstract:
为探究海马齿高盐环境耐受性的分子机制，根据海马齿cdna文库序列设计引物，采用race（rapidamplificationofcdnaends）技术克隆得到海马齿小分子热激蛋白sphsp18.1基因序列.blast分析表明，该cdna序列（968bp）含完整开放阅读框（orf）全长为489bp，编码162个氨基酸.等电点（pi）为6.19，分子量（mr）大小为18605.90.sphsp18.1编码氨基酸序列具有分子伴侣蛋白特有的acd保守区域（alphacrystallindomain），与拟南芥、西瓜等植物的细胞质i型小热激蛋白具较高同源性（>75%）.将sphsp18.1基因克隆到pucm-t原核表达载体上，得到重组菌株，通过过量表达探究其耐热胁迫能力.结果表明，在37℃下重组菌株与野生型菌株的生长曲线基本相似，但在高温（45℃）下，重组菌株较对照组菌株存活率有显著提高.荧光实时定量pcr分析表明，在不同浓度海水培养条件下，海马齿根部均表达sphsp18.1基因，且随着海水浓度的增加，该基因的表达量呈现上升的趋势.说明sphsp18.1基因在海马齿应答高盐胁迫过程中起到一定的作用.
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