从银叶真藓(Bryum argenteum)转录组数据库出发,使用Pfam数据库提供的HMM模型共得到33条长度大于200aa,注释的热休克蛋白BaHSP70;其中2条(BaHSP70-1,BaHSP70-2)具有完整ORF,NCBI核酸数据库登录号为KP087877和KP087878。使用生物信息学在线分析工具和软件,对真藓HSP70的两条蛋白质序列从氨基酸组成、保守结构域、理化性质、疏水性/亲水性、信号肽、蛋白质结构、模体的识别及同源性分析等方面进行了预测和分析。结果表明:2条BaHSP70s基因序列ORF全长分别为2396 bp和2356 bp,分别编码649aa和650aa。序列模体分析表明BaHSP70s和其它报道的植物HSP70均含有4个相同的模体,并且各模体在蛋白质序列上顺序一致。通过对2条BaHSP70s进行氨基酸多序列比对及基因树分析,发现BaHSP70-1和BaHSP70-2雪莲相似度最高,分别是91.2%和86.6%。本研究为进一步研究HSP70基因的克隆和功能验证奠定了基础。
Our study utilizes the RNA-seq technology which is based on the Hi-seq 2000 sequencing platform from BGI to acquire the RNA sequences data from Bryum argenteum dealt with dehydration and rehydration stress. Using the Hidden Markov Model provided by the Pfam database,we have identified 33 HSP70 sequences with the length longer than 200 aa. Two of them(BaHSP70-1,BaHSP70-2) contain Open Reading Frame,the Accession No. in NCBI nucleotide database are KP087877 and KP087878,respectively. Applying web-based bioinformatics tools and softwares,we predicted and analyzed B. argenteum Heat Shock Protein 70 sequences from the following aspects:amino acid component, conserved domain, physicochemical property, hydrophobicity and hydrophily, singal peptide,protein structure,motif recognition,homologous analysis. The result showed the length of two possessed complete ORF HSP70 s are 649 aa and 650 aa respectively. The length of corresponding anscripts in the transcriptome database are 2 396 bp and 2 356 bp. Sequence motif analysis indicates B. argenteum and other plant species have 4identical motifs. Meanwhile,the order of the motifs arranged in protein sequence is the same in all selected species.Multiple sequence alignment and homologous analysis demonstrated that HSP70-1 and HSP70-2 are most similar to Saussurea up to the similarity of 91. 2% and 86. 6% respectively. This study laid fundation for the B. argenteum further research in gene clone and functional confirmation.