杜梨类钙调磷酸酶B亚基蛋白PbCBL10基因的克隆和表达特性研究Cloning and expression characteristics of a Calcineurin B-like protein gene(PbCBL10) from Pyrus betulaefolia
李慧,丛郁,常有宏,蔺经,盛宝龙
摘要(Abstract)：
【目的】为了探明杜梨类钙调磷酸酶B亚基蛋白(Calcineurin B-like protein,CBL)基因的序列特征和表达特点,【方法】以杜梨幼苗为试材,运用同源克隆和半定量RT-PCR对PbCBL10基因进行克隆和表达分析。【结果】结果表明,PbCBL10基因编码区cDNA长度为801 bp,编码的多肽由266个氨基酸组成。该多肽预测的等电点为4.56,估计的相对分子质量为30.45 ku。其对应基因组DNA序列长1 983 bp,包括9个外显子和8个内含子。通过PSORT进行亚细胞定位分析发现PbCBL10蛋白位于质膜上的几率最大。PbCBL10基因编码的多肽具有4个钙离子结合域EF手形结构和1个钙调磷酸酶A亚基结合位点。PbCBL10与番茄SlCBL10(NP_001239045)和拟南芥AtCBL10(NP_195026)蛋白间的同源性较高,分别为82%和77%,并与AtCBL10亲缘关系最近。PbCBL10基因在杜梨幼苗根、茎和叶片中均为诱导型表达,50~200 mmol.L-1CaCl2、20μmol.L-1ABA、100 mmol.L-1NaCl、10%(w/v)PEG6000或180 mmol.L-1甘露醇处理后其表达量明显上调。【结论】PbCBL10基因具备植物CBL基因家族的固有特征,能够响应胞内钙浓度变化,对ABA、盐碱、干旱和渗透胁迫均存在转录响应。
关键词(KeyWords)： 杜梨;类钙调磷酸酶B亚基蛋白;基因克隆;表达特性
基金项目(Foundation): 国家自然科学基金(31101529);; 江苏省农业科技自主创新资金(CX(11)4050)
作者(Author): 李慧,丛郁,常有宏,蔺经,盛宝龙
DOI: 10.13925/j.cnki.gsxb.2012.04.013
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