松材线虫和拟松材线虫14-3-3蛋白基因全长cDNA克隆与分析

目的]对松材线虫和拟松材线虫的14-3-3蛋白基因进行全长cDNA克隆，并进行序列分析。方法]根据GenBank数据库中14-3-3蛋白EST序列BXC56（登录号为FG589472）、BMCl20（登录号为FG589351），采用RACE技术获取松材线虫和拟松材线虫14-3-3蛋白的全长cDNA克隆Bxl4-3-3a和Bml4-3-3a，并对克隆片段序列进行分析。结果]Bx14-3-3a全长1039bp，包含一个756bp的开放阅读框，编码蛋白含有251个氨基酸残基，包含2个14-3-3蛋白标签，编码蛋白分子量为61．43kD，等电点为4．88。Bx14-3-3a基因组结构中包含4个外显子和3个内舍子序列。Bm14-3-3a全长992bp。有与Bx14-3-3a编码完全相同的氨基酸序列。序列比对结果表明，14-3-3在真核生物间高度保守，Bx14-3-3A和Bm14-3-3A蛋白在系统进化上与线形动物门（Nematomorpha）的物种近缘关系较近，与植物线虫南方根结线虫（Meloidogyneincognita）系统发育关系上最为亲近。结论]Bx14-3-3a和Bm14-3-3a的成功克隆，为进一步研究其功能及其调控因子对形态、致病力、繁殖力和环境适应性等方面的分子调控机制奠定了重要基础。

Full-length cDNA Cloning and Sequences Analysis of 14-3-3 Genes Generated from Bursaphelenchus xylophilus and B. mucronatus

Objective] The study aimed to clone the full-length cDNA of 14-3-3 gene from B.xylophilus and B.mucronatus,respectively,and sequence analysis was carried out.Method] Based on the EST BXC56(Accession No.FG589742) and BMC120(Accession No.FG589351),the full length cDNA of Bx14-3-3a and Bm14-3-3a was cloned by the method of RACE and the bioinformatical analysis of the cDNAs was carried out.Result] Bx14-3-3a was 1 039 bp long and contained a 756 bp-long open reading frame(ORF).The deduced protein Bx14-3-3A contained 251 amino acids with molecular weight of 61.43 ku and PI of 4.88 and two 14-3-3 protein family signatures.Bm14-3-3a was 992 bp long and the deduced protein Bm14-3-3A was same to Bx14-3-3A.Alignment and phylogeny analysis showed that Bx14-3-3A and Bm14-3-3A had close relationship with the 14-3-3 proteins from Nematomopha species,especially closer to the Meloidogyne incognita.Conclusion] The full-length cDNA of Bx14-3-3a and Bm14-3-3a from B.xylophilus and B.mucronatus was successfully cloned,which provide important base for their function,and the molecular regulation mechanism to the morphological character,pathologinity,reproduction and environmental adaptability as a regulation factors.