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OALib Journal期刊
ISSN: 2333-9721
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甘蔗真核生物翻译起始因子5a基因的克隆与表达分析*
DOI: 10.3724/SP.J.1145.2015.04008, PP. 1-11
Keywords: 甘蔗,真核生物翻译起始因子5a,甘蔗黑穗病菌,生物信息学,实时荧光定量pcr
Abstract:
真核生物翻译起始因子5a（eukaryotictranslationinitiationfactor5a，eif5a）是一种在动植物和真菌体内普遍存在的蛋白质。本研究首先从黑穗病菌胁迫下甘蔗ssh(suppressionsubtractivehybridization，抑制消减杂交)文库中获得一条与玉米eif5a基因（genbankaccessionnumber：nm_001112136.1）同源性为93%的甘蔗est序列；其次，以此序列作为探针，通过电子克隆技术获得一条甘蔗eif5a基因的cdna拼接序列；最后，经rt-pcr扩增和测序验证，结果显示电子克隆序列与测序序列一致，将该序列命名为sceif5a（genbankaccessionnumber：kj577595）。生物信息学分析显示，sceif5a基因cdna全长1,174bp，含有长度为483bp，编码160个氨基酸的完整开放读码框；sceif5a蛋白是酸性稳定蛋白，分子量为17453.6da，含有12个保守氨基酸序列，推测定位于细胞质。rt-qpcr（real-timequantitativepcr，实时荧光定量pcr）分析结果表明，在黑穗病菌、水杨酸、茉莉酸甲酯、脱落酸胁迫下，sceif5a均上调表达，推测sceif5a基因在甘蔗响应黑穗病菌侵染过程中被诱导表达，且其表达受内源激素信号通路的调控。研究结果为研究sceif5a基因在甘蔗与黑穗病菌互作中的作用积累了基础数据，并为进一步深入鉴定该基因的功能奠定一定的理论和实践基础。
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