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OALib Journal期刊
ISSN: 2333-9721
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龙眼胚性愈伤组织cu/zn-sod分子伴侣基因ccs的克隆及其在体胚发生过程中的表达分析
DOI: 10.3724/SP.J.1145.2012.00351, PP. 351-358
Keywords: 龙眼,体细胞胚胎发生,ccs,克隆,生物信息学,实时荧光定量pcr
Abstract:
为了解cu/zn超氧化物歧化酶分子伴侣基因（ccs）在龙眼体胚发生过程中的表达调控机制，以龙眼松散型胚性愈伤组织为材料，采用rt-pcr法，克隆了长960bp含有完整开放阅读框的龙眼dlccs核酸序列，其编码一个含有319个氨基酸的蛋白质（genbank登录号：fj973472）.生物信息学分析显示：该蛋白为亲水的、稳定的、含有跨膜结构域的酸性蛋白质，定位于叶绿体；与毛果杨、葡萄、拟南芥、水稻和锦鸡儿具有较高同源性，含有植物ccs蛋白所特有的3个保守结构域；预测其通过不同的磷酸化方式，改变酶活性及蛋白构象，参与龙眼体胚中超氧化物代谢以及金属离子转运等生物学过程，在氧化还原过程中发挥作用.利用实时荧光定量pcr（qpcr）技术研究该基因在龙眼体胚发生发育过程中的表达情况，结果显示，从松散型胚性愈伤组织（stage1）到胚性紧实球形结构阶段（stage4），dlccsmrna的转录水平较低且波动小；当胚性紧实球形结构进一步发育到子叶形胚阶段（stage8），其表达量迅速增加，在成熟胚阶段（stage9）达到最高峰，提示dlccs可能在龙眼体胚的中晚期起主要作用.
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