论文部分内容阅读
根据已知EST片段结合RT-PCR和RACE技术,从东方山羊豆(Galega Orientalis)中克隆到一个水通道蛋白(AQP)基因,命名为GoAQP(GenBank登录号:HM 803185)。测序和生物信息学分析表明,此序列全长1258bp,开放读码框(ORF)870 bp,编码289个氨基酸。GoAQP包含MIP家族信号序列HINPAVT/SFG,高等植物PIP高度保守序列GGANXXXXGY和TGI/TNPARSL/FGAAI/VI/VF/YN。与拟南芥AQP基因家族的系统进化分析表明,GoAQP属于PIP1亚型AQP蛋白。Real-Time PCR检测结果表明,GoAQP基因在根中表达量最高,茎中表达量最少;且受NaCl和PEG胁迫表达上调,NaCl表达呈双峰分布。因此GoAQP基因可能参与了东方山羊豆耐盐性调控。同时成功构建超表达载体pCAMBIA-1302-GoAQP,为转基因验证基因功能创造了基础。
AQP gene was cloned from Galega Orientalis and named GoAQP (GenBank accession number: HM 803185) based on the known EST fragments combined with RT-PCR and RACE techniques. Sequencing and bioinformatics analysis showed that this sequence was 1258bp in length and 870 bp in open reading frame (ORF) encoding 289 amino acids. GoAQP contains the MIP family signal sequence HINPAVT / SFG, the higher plant PIP highly conserved sequences GGANXXXXGY and TGI / TNPARSL / FGAAI / VI / VF / YN. Phylogenetic analysis of the AQP gene family in Arabidopsis showed that GoAQP belongs to PIP1 subtype AQP protein. Real-Time PCR results showed that GoAQP gene had the highest level of expression in roots and the lowest in stems. The expression of GoAQP gene was up-regulated by NaCl and PEG, and the expression of NaCl was bimodal. So GoAQP gene may be involved in the regulation of salt tolerance of goat Beans. At the same time, we successfully constructed the overexpression vector pCAMBIA-1302-GoAQP, which laid the foundation for the function of transgenic gene.