以‘南林895’杨(Populus deltoides×P.euramericana cv ‘Nanlin895’)为材料,通过RACE-PCR技术,获得了NAC1基因的全长cDNA,命名为PdNAC1。该序列含有1个长906 bp的开放阅读框,编码302个氨基酸。该蛋白的分子质量为34.23 ku,等电点为7.44。分析其氨基酸序列发现,该蛋白具有典型NAC转录因子特征,NAM结构域位于11～135个氨基酸之间,含有A、B、C、D、E 5个亚结构域,该区域可以结合DNA; 而C端含有一个高度变异的转录激活区。利用杨树原生质体对PdNAC1蛋白进行细胞亚定位,结果发现PdNAC1不仅明显定位于细胞核中,而且在细胞膜上也有表达。但PdNAC1是否具有相应的跨膜结构域尚不明确。
Abstract
NAC transcription factors are the key transcription regulatory factors which involved in plant multiple biological functions, such as plant development, hormones regulation, and environmental stimuli. The full length cDNA sequence of a NAC-like gene from the hybrid of Populus deltoides × P. euramericana cv ‘Nanlin895’, was cloned by RT-PCR and RACE methods, named PdNAC1, which contains an ORF of 906 bp and encodes 302 amino acids with the molecular weight of about 34.23 ku ond pI of 7.44. The putative protein sequence analysis showed that PdNAC1contained conserved NAC domain including five sub-domains A, B, C, D, and E which can bind DNA. Meanwhile, the C-terminal of PdNAC1 is highly different and that is very typical in NAC transcription factors. Furthermore, the results of sub-cellular localization in poplar protoplast showed that the PdNAC1 transcription factor is mainly located in the cell nucleus, but also translocated on the cell membrane. However,further investigation would be needed to reveal whether the PdNAC1 is corresponding to transmembrane domain.
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基金
收稿日期:2014-04-22 修回日期:2014-06-20
基金项目:国家林业公益性行业科研专项项目(201304111); 国家自然科学基金项目(31200507); 江苏省基础研究计划(自然科学基金)-面上研究项目(BK2012817); 江苏高校优势学科建设工程资助项目(PAPD)
第一作者:王浩然,硕士生。*通信作者:陈英,副教授。E-mail: ychen@njfu.edu.cn。
引文格式:王浩然,邵志龙,朱燕宇,等. ‘南林895’杨PdNAC1基因克隆及蛋白的亚细胞定位[J]. 南京林业大学学报:自然科学版,2015,39(3):50-54.
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