CAJ | 학술논문

为了研究苯丙氨酸解氨酶基因与大蕉(Musa ABB cv.Dongguandajiao)抗枯萎病的关系,利用RT-PCR和RACE技术克隆了大蕉苯丙氨酸解氨酶基因全长cDNA.此cDNA长1 300 bp,包含一个长为1 191 bp,编码397个氨基酸的完整开放阅读框(ORF),推导的氨基酸序列与水稻PAL基因氨基酸序列同源性达89%,将此基因命名为M-PAL.Southern杂交结果表明大蕉中存在一个包含4-5个PAL基因的基因家族,将此基因克隆到大肠杆菌表达载体pET32(a+)中,表达的蛋白质分子量大小与推导的相一致,并且表达的蛋白质表现出PAL酶活性.对接种香蕉枯萎病菌4号生理小种(Fusarium oxysporum f.sp.cubense(FOC)race4)后大蕉叶片中M-PAL基因的转录谱进行研究表明,在接种枯萎病菌后,M-PAL基因在叶片中的转录水平提高,因此推测M-PAL基因的表达可能与香蕉枯萎病抗性相关.
위료연구분병안산해안매기인여대초(Musa ABB cv.Dongguandajiao)항고위병적관계,이용RT-PCR화RACE기술극륭료대초분병안산해안매기인전장cDNA.차cDNA장1 300 bp,포함일개장위1 191 bp,편마397개안기산적완정개방열독광(ORF),추도적안기산서렬여수도PAL기인안기산서렬동원성체89%,장차기인명명위M-PAL.Southern잡교결과표명대초중존재일개포함4-5개PAL기인적기인가족,장차기인극륭도대장간균표체재체pET32(a+)중,표체적단백질분자량대소여추도적상일치,병차표체적단백질표현출PAL매활성.대접충향초고위병균4호생리소충(Fusarium oxysporum f.sp.cubense(FOC)race4)후대초협편중M-PAL기인적전록보진행연구표명,재접충고위병균후,M-PAL기인재협편중적전록수평제고,인차추측M-PAL기인적표체가능여향초고위병항성상관.
A full-length cDNA encoding PAL was cloned from leaves of plantain (Musa ABBcv. Dongguandajiao)by RT-PCR and RACE. The cDNA was 1 300 bp in length and contained a complete ORF of 1 191 bp which encodes a protein of 397 amino acid residues. Its deduced amino acid sequence had 89% identity with rice PAL and then this gene was hence designated as M-PAL. Southern hybridization analysis demonstrated that a small family of four to five PAL genes exists in plantain banana. The complete coding sequence of M-PAL was cloned into an expression vector pET32 (a+) and a protein about 43 KD expressed in Escherichia coli BL21 (DE3) was the same as deduced molecular weight and showed phenylalanine ammonia-lyase activity. The transcript of M-PAL gene in the leaves was increased when the plantlets of plantain were inoculated with Fusarium oxysporun f. sp.cubense, indicating that the expression of M-PAL may be related to resistance of banana Fusarium wilt.