玉米氨基酸转运蛋白ZmAAP基因的克隆及序列分析

Cloning and Sequence Analysis of Amino Acid Transporter ZmAAP Gene in Maize

克隆得到玉米中AAP基因的核酸序列,通过对其编码蛋白理化性质、跨膜区、高级结构等方面进行生物信息学分析,为进一步深入研究玉米AAP基因的功能提供理论依据。采用同源克隆技术得到含有完整编码阅读框的一段玉米未知功能基因,使用ProtParam、ProtScale、TargetP v1.01 server、SignalP4.1 server、TMHMM services v.2.0、SOPMA、Phyre2、DNAman等软件对该基因进行生物信息学分析。结果表明:克隆的玉米未知功能基因全长cDNA序列为1396 bp,包括1212 bp的开放阅读框,编码的氨基酸数403个,分子质量42.88 kD,理论等电点9.24,正、负电荷残基总数分别是18和30,在组成蛋白编码的20种氨基酸中丝氨酸所占比例最高,为13.4%,而组氨酸比例最低,为1.0%。AAP蛋白包含信号肽,剪切位点位于38、39残基处;编码区核苷酸序列与其他作物对比后发现,与大麦AAP1基因的相似性最高,相似度为87%。通过生物信息学分析初步证明从玉米中克隆的基因为AAP的核酸序列,命名为ZmAAP。

The nucleic acid sequence of AAP gene in maize was obtained by cloning,and bioinformatics analysis was carried out on physicochemical properties,transmembrane region and high-level structure of the encoded protein,so as to provide theoretical basis for further study on the function of AAP gene in maize.Homologous cloning technology was used to obtain an unknown functional gene with complete coding reading frame,and bioinformatics analysis of the gene was carried out by using ProtParam,ProtScale,TargetP v1.01 server,SignalP4.1 server,TMHMM services v.2.0,SOPMA,Phyre2,DNAman and other softwares.The results showed that the full-length cDNA sequence of the unknown functional gene was 1396 bp,including an open reading frame of 1212 bp,encoding 403 amino acids,its molecular mass was 42.88 kD,its theoretical isoelectric point was 9.24,and the total number of positive and negative charge residues were 18 and 30,respectively.Among the 20 amino acids encoded by the protein,serine accounted for the highest proportion(13.4%),while histidine accounted for the lowest proportion(1.0%).AAP protein contains signal peptide,and its splice site is located at residues 38 and 39.Compared with other crops,the nucleotide sequence of maize coding region has the highest similarity with barley AAP1 gene,with a similarity of 87%.Through bioinformatics analysis,the gene cloned from maize is preliminarily proved to be the nucleic acid sequence of AAP,named ZmAAP.