草菇MAPKKK同源基因的克隆及序列分析  

Cloning and Sequence Analysis of Mitogen-activated Protein Kinase Kinase Kinase Homolog from Volvariella volvacea(VV-MAPKKK)

作　　者：汪虹[1] 鲍大鹏[1] 陈明杰[1] 陈辉[1] 冯爱萍[1] 

机构地区：[1]国家食用菌工程技术研究中心农业部应用真菌资源与利用重点开放实验室上海市农业遗传育种重点开放实验室上海市农业科学院食用菌研究所,上海201106

出　　处：《微生物学通报》2010年第11期1606-1609,共4页Microbiology China

基　　金：上海农科院2008年度院发基金[No.上海农科院农科发2008(16)];上海科技兴农重点攻关项目(No.沪农科攻字2007第6-2号);农业部公益性行业科研专项项目(No.nyhyzx07-008);上海市浦江人才计划项目(No.08PJ14087)

摘　　要：根据担子菌丝裂原活化蛋白质激酶激酶激酶(MAPKKK)蛋白的保守序列设计两对简并引物,通过巢式简并PCR方法获得草菇VV-MAPKKK基因中的保守片段,然后通过和草菇基因组信息比对,获得了VV-MAPKKK基因全长序列。VV-MAPKKK基因长度为4434bp,包含4个内含子,编码1405个氨基酸残基,推定的氨基酸序列与新型隐球菌(Cryptococcus neoformans)、巴西芽生菌(Paracoccidioides brasiliensis)和异旋孢腔菌(Cochliobolus heterostrophus)的MAPKKK同源蛋白相似性分别为58%、57%和56%。对VV-MAPKKK蛋白的系统发生学分析的结果表明,VV-MAPKKK与担子菌中的Hog信号传导途径的MAPKKK同源蛋白聚在同一进化支上,这些数据都支持所获得的VV-MAPKKK为Hog-MAPKKK蛋白在草菇中的同源物的推定。Two pairs of degenerate primers were designed according to the conserved amino acid sequences of several mitogen-activated protein kinase kinase kinases（MAPKKK） from basidiomycetes.Based on the conserved DNA sequence,which was obtained by nested degenerate PCR,a whole DNA sequence of MAPKKK of Volvariella volvacea（VV-MAPKKK） was obtained by blasting with Volvariella volvacea genome.VV-MAPKKK gene,containing 4 introns and codeing for a 1405 aa protein（VV-MAPKKK）,is 4434 bp in length.The similarity of VV-MAPKKK compared with Hog-MAPKKK protein from Cryptococcus neoformans,Paracoccidioides brasiliensis and Cochliobolus heterostrophus is 58%,57% and 56%,respectively.Phylogenetic analysis showed that VV-MAPKKK and Hog-MAPKKK protein homolog belong to the same clade.These data supported that VV-MAPKKK protein is a Hog-MAPKKK protein homolog from Volvariella volvacea.

关 键 词：草菇 丝裂原活化蛋白质激酶激酶激酶(MAPKKK) 克隆 序列分析 

分 类 号：Q943.2[生物学—植物学] S646.13[农业科学—蔬菜学]