草地早熟禾硝酸盐转运蛋白NRT1/PTR FAMILY 5.8基因的克隆及生物信息学分析

硝态氮是植物主要的氮素营养来源,与植物的生长发育与抗性能力密切相关。硝态氮的吸收与运转对提高植物氮素利用效率有调控作用,其中硝酸盐转运蛋白发挥重要作用。本研究通过RT-PCR的方法克隆得到草地早熟禾硝酸盐转运蛋白NRT1/PTR FAMILY 5.8基因的编码区序列,并进行生物信息学分析。结果表明,该基因编码区长度为1434 bp,共编码477个氨基酸。NRT1/PTR FAMILY 5.8蛋白分子量为51.7 kD,等电点为4.9。NRT1/PTR FAMILY 5.8蛋白共有8个跨膜结构域,3个糖基化,位点无信号肽,预测定位于质膜或内质网中。NRT1/PTR FAMILY 5.8蛋白含有结构域PTR2属于MFS蛋白超家族。草地早熟禾NRT1/PTR FAMILY 5.8蛋白与二穗短柄草,粗山羊草的相似度较高,而且不同物种之间的NRT1/PTR FAMILY 5.8与NRT1/PTR FAMILY 5.9的同源进化关系较近。本研究为进一步研究NRT1/PTR FAMILY 5.8基因的功能及硝酸盐转运途径具有重要意义。

Cloning and Bioinfomatic Analysis of Nitrate Transporters NRT1/PTR FAMILY 5.8 Gene in Kentucky Bluegrass

Nitrate nitrogen,which is the main source of nitrogen nutrition in plants,is closely related to plant growth,development and resistance.The absorption and transportation of nitrate nitrogen play a regulatory role in improving nitrogen-use efficiency in plants,in which nitrate transporters play an key role.In this study,the coding sequence of nitrate transporter NRT1/PTR FAMILY 5.8 gene of Kentucky bluegrass was cloned using RT-PCR and analyzed by bioinformatics.The results demonstrated that the length of coding region of the NRT1/PTR FAM-%ILY 5.8 gene was 1434 bp,encoding 477 amino acids.The analysis of physicochemical properties predicted that the molecular weight of the protein was 51.7 kD,the isoelectric point was 4.9,and the content of Leu was the highest,accounting for 10.5%.The NRT1/PTR FAMILY 5.8 protein had 8 transmembrane domains,no signal peptide and 3 glycosylation sites,which may be located in the plasma membrane or endoplasmic reticulum.The NRT1/PTR FAMILY 5.8 protein contained a PTR2 domain,which belonged to the major facilitator superfamily(MFS).The secondary structure was mainly composed ofα-helix and random curl.Phylogenetic analysis showed that the sequence of NRT1/PTR FAMILY 5.8 protein of Kentucky bluegrass was highly similar to that of Brachypodium distachyon and Aegilops tauschii,and the homologous evolutionary relationship between NRT1/PTR FAMILY 5.8 and NRT1/PTR FAMILY 5.9 among different species was similar.Cloning and sequence analysis of the NRT1/PTR FAMILY 5.8 gene of Kentucky bluegrass are of great significance to further study on the function of this gene and the nitrate transport pathway of Kentucky bluegrass.