摘要
利用来自Forest TreeDB数据库中热胁迫cDNA文库ESTs聚类、拼接组装的Unigene以及基因GO注释,得到了假定的火炬松DREB1基因,全长1 293bp,具有完整的蛋白编码框的cDNA序列。采用DNAMAN软件分析碱基组成、限制酶切位点和重复序列等,结果发现,该序列与其他物种中克隆得到的DREB1具有较高的同源性,初步断定所电子克隆得到的cDNA序列为火炬松DREB1基因(PteaDREB1)。在此基础上,利用网上的公共数据库和相关软件(Antheprot等)对PteaDREB1编码的理化性质和一级结构进行分析,并模拟了其二级结构和三级结构。结果表明,该蛋白为疏水性非球形可溶蛋白,含有295个氨基酸,分子量为32.4kD,等电点为8.22;其二级结构以螺旋和卷曲为主,三级结构具有DREB蛋白家族中典型的结合DNA的AP2结构域。这进一步说明所克隆到的cDNA片断为火炬松DREB1基因。上述研究结果可为PteaDREB1基因下一步的分子克隆、功能鉴定和应用奠定基础,具有一定的现实意义和应用前景。
Based on the results of gene assembly and gene annotation of ESTs from heat induced cDNA library in the ForestTreeDB through program CAP3 and software AmiGo,putative Pinus DREB1 gene of a 1293 bp-length cDNA sequence with an full ORF was obtained.The basic composition,the restriction enzyme sites and the repeat fragments of PteaDREB1 gene were analyzed.It was found that PteaDREB1 gene encoded 421 amino acids with MW 32.4 kD and PI 8.22.The gene shared high similarity with other plant DREB1 genes,indicating that PteaDREB1 might be a new member of DREB1 family.On this base,through public database such as CPHmodel and related softwares,the primary structure of the protein PteaDREB1 such as physical and chemical characterization was analyzed,and then the secondary and tertiary structure was emulated.The results showed that the protein encoded by PteaDREB1 might be hydrophilic and not a globular with some flexible domains,its N-terminus presented a basic amino acid-rich nuclear localization signal.The secondary structure are mainly Helix and Coli and part of which tertiary structure containeda classical DNA-combining domain AP2,similar to DREB proteins family.The above results may play an important role in the further research for isolatiion,identification and application of PteaDREB1 gene.
出处
《生物信息学》
2010年第1期43-46,共4页
Chinese Journal of Bioinformatics
基金
广东省自然科学基金重点项目(7118123)
