甘蓝型油菜组蛋白乙酰转移酶Bn-HAM2基因的克隆及生物信息学分析

Cloning and Bioinformatics Analysis of Histone Acetyltransferase BnHAM2 Gene from Rapeseed(Brassica napus L.)

HAM2基因在植物生长、发育和应激反应中起重要的调控作用。本研究以油菜品种‘美国油王88’和‘南油868’为材料,通过PCR技术扩增HAM2完整编码区,利用生物信息软件进行序列分析,并通过PlantCARE对其启动子序列进行分析。结果表明,Bn-HAM2-MY88和Bn-HAM2-NY868的开放阅读框长度均为1326 bp,编码441个氨基酸,分子量分别约为50.81 kD和50.86 kD,理论等电点分别为7.88和7.56;亚细胞定位预测分析发现该基因编码的蛋白主要分布于细胞核;功能结构域分析结果证实,Bn-HAM2-MY88和Bn-HAM2-NY868蛋白都包含有保守的MYST结构域,属于MYST亚家族成员;多序列比对及进化树分析结果表明,Bn-HAM2-MY88和Bn-HAM2-NY868与NCBI中公布的甘蓝型油菜组蛋白乙酰转移酶氨基酸序列(XP_013715766)相似性最高,分别为99.77%和100%,与拟南芥组蛋白乙酰转移酶序列(KAG7554717.1)相似性高达90%以上;启动子分析表明,除含有真核生物启动子TATA和CAAT固有元件外,还存在W-box、P-box、TGA-element、TC-rich repeats等多种响应逆境胁迫的顺式元件。本研究结果为进一步研究Bn-HAM2-MY88和Bn-HAM2-NY868的生物学功能提供了基础资料。

HAM2 gene plays an important regulatory role in plant growth,development and stress response.In this study,the open reading frame(ORF),sequences of two Bn-HAM2 genes were cloned respectively from Brassica napus cultivar’American Youwang88’and’Nanyou868’by PCR,and their sequences were analyzed by bioinformatics.The promoter region was also analyzed by PlantCARE.The results showed that both of the BnHAM2-MY88 and Bn-HAM2-NY868 contained 1326 bp ORFs encoding proteins of 441 amino acids,with the molecular weights of about 50.81 kD and 50.86 kD,and the theoretical isoelectric points of 7.88 and 7.56,respectively;Subcellular localization prediction analysis demonstrated that the Bn-HAM2-MY88 and Bn-HAM2-NY868mainly distributed in nucleus.Functional domain analysis confirmed that both proteins contained conserved MYST domains and belonged to the MYST subfamily;The results of multiple sequence alignment and phylogenetic tree analysis showed thatBn-HAM2-MY88 and Bn-HAM2-NY868 shared high similarity(99.77%and 100%respectively)with the histone acetyltransferase amino acid sequence of Brassica napus(XP_013715766),and more than90%with the histone acetyltransferase sequence of Arabidopsis(KAG7554717.1);Promoter analysis showed that,besides the eukaryotic promoter TATA and CAAT natural elements,the promoter sequence of Bn-HAM2 also had cis elements in response to stress,such as W-box,P-box,TGA-element and TC-rich repeats,etc..This study provides basic data for further studying the biological function of Bn-HAM2-MY88 and Bn-HAM2-NY868.