CAJ | 학술논문

根据R基因及其调控基因保守序列设计简并性引物,对白粉菌接种和未接种处理的一对抗病和感病的小麦-黑麦等位突变易位系TAM104R和TAM104S总RNA进行RT-PCR扩增,得到一个诱导表达的cDNA片段.序列分析表明,该片段全长2 474 bp,其中含有一个822 bp的完整开放阅读框,推测其编码一个有273个氨基酸残基、分子量约31 kD的蛋白质分子.蛋白质的氨基酸序列比对显示,该蛋白质分子具有C2HC锌结合motif CX2CX4HX4C结构和锌指domain,可见克隆的cDNA是一个锌指蛋白基因,命名为TaZF.Southern杂交表明,TaZF在抗病易位系TAM104R的基因组中是多拷贝的.半定量RT-PCR分析显示,TaZF基因属组成型表达、但受白粉菌诱导表达上调的基因,推测其与白粉病菌的侵染过程相关.基因组DNA专化扩增、克隆和测序揭示TaZF基因无内含子.
근거R기인급기조공기인보수서렬설계간병성인물,대백분균접충화미접충처리적일대항병화감병적소맥-흑맥등위돌변역위계TAM104R화TAM104S총RNA진행RT-PCR확증,득도일개유도표체적cDNA편단.서렬분석표명,해편단전장2 474 bp,기중함유일개822 bp적완정개방열독광,추측기편마일개유273개안기산잔기、분자량약31 kD적단백질분자.단백질적안기산서렬비대현시,해단백질분자구유C2HC자결합motif CX2CX4HX4C결구화자지domain,가견극륭적cDNA시일개자지단백기인,명명위TaZF.Southern잡교표명,TaZF재항병역위계TAM104R적기인조중시다고패적.반정량RT-PCR분석현시,TaZF기인속조성형표체、단수백분균유도표체상조적기인,추측기여백분병균적침염과정상관.기인조DNA전화확증、극륭화측서게시TaZF기인무내함자.
Reverse Transcription-Polymerase Chain Reaction(RT-PCR) was performed on total RNA of resistant-line TAM104R and susceptible-line TAM104S using 33 primers designed according to conservative domain of plant R-genes and the sequence of barley powdery mildew resistance genes Mlo,Mla1,Mla6 and wheat leaf rust resistance gene Lrk10.A polymorphic cDNA fragment TaZF was cloned and sequenced.The Open Reading Frame (ORF) of TaZF is comprised of 822 base pairs which encodes a zinc finger-like DNA or RNA-binding protein with 273 amino acids and molecular weight of 31 kD.TaZF is more than one copy gene in TAM104R genome based on southern blot.It is constitutively expressed,but level of expression was enhanced in tissue infected by Erysiphe Graminis.There is no intron in TaZF genome DNA.