CAJ | 학술논문

14-3-3蛋白是一类在真核生物组织中广泛表达并主要通过与不同的靶蛋白互作参与一系列细胞信号传导和代谢过程的酸性类调控蛋白。通过对辣椒（ Capsicum annuum L．）均一化cDNA文库的筛选，首次分离获得一个辣椒推定蛋白14-3-3的全长cDNA，长度为1251 bp，含有一个长度为765 bp的开放阅读框（ open reading frame，ORF），其推定的蛋白质含有254个氨基酸残基，理论分子量约为28．7 ku，等电点为4．8，无明显信号肽及跨膜区，定名为Ca14-3-3a。序列分析显示Ca14-3-3a属于非ε样14-3-3蛋白，与番茄TFT10及烟草T14-3g的氨基酸序列一致性分别高达96％和93％，尤其是介导14-3-3蛋白与SPS（ Sucrose-6-Phosphate Synthase ）互作的C端序列完全一致，暗示其可能参与辣椒蔗糖代谢。实时定量PCR结果显示Ca14-3-3a基因在辣椒青枯菌处理下表达水平持续下调，推测其还可能通过与不同的靶蛋白结合参与植物病原菌防御应答。
14-3-3단백시일류재진핵생물조직중엄범표체병주요통과여불동적파단백호작삼여일계렬세포신호전도화대사과정적산성류조공단백。통과대랄초（ Capsicum annuum L．）균일화cDNA문고적사선，수차분리획득일개랄초추정단백14-3-3적전장cDNA，장도위1251 bp，함유일개장도위765 bp적개방열독광（ open reading frame，ORF），기추정적단백질함유254개안기산잔기，이론분자량약위28．7 ku，등전점위4．8，무명현신호태급과막구，정명위Ca14-3-3a。서렬분석현시Ca14-3-3a속우비ε양14-3-3단백，여번가TFT10급연초T14-3g적안기산서렬일치성분별고체96％화93％，우기시개도14-3-3단백여SPS（ Sucrose-6-Phosphate Synthase ）호작적C단서렬완전일치，암시기가능삼여랄초자당대사。실시정량PCR결과현시Ca14-3-3a기인재랄초청고균처리하표체수평지속하조，추측기환가능통과여불동적파단백결합삼여식물병원균방어응답。
14-3-3 is a family of acidic proteins expressing widely in various organs or tissues in all eu-karyotes and acting as an important regulator in a vast array of signaling cascades and metabolic processes by interacting with their targeting proteins .The members in the 14-3-3 family can be subdivided into εgroup and non-εgroup,the majority members in this family even in the model plants Arabidopsis and rice remain to be elucidate.Herein,a full length cDNA of a putative member of the 14-3-3 family,designated as Ca14-3-3a, was isolated from a normalized cDNA library of Capsicum annuum.It is 1 251 bp in length ,harboring an open reading frame of 765 bp in length,encoding a protein with 254 aa in length.Its theoretical molecular mass is 28.7D,its calculated isoelectric point is 4.8,and no signal peptide and transmembrane domain were observed in this putative protein.By homologous alignment analysis ,the putative protein exhibited a 96%and 93%iden-tity with TFT10 of tomato and T14-3g of tobacco in amino acids sequence ,suggesting that it is a protein of 14-3-3 belonging to ε group other than non-εgroup.Especially,the C-terminus sequences of 14-3-3,impor-tant for the interaction of 14-3-3s with SPS,are high similarity between TFT10,T14-3g and Ca14-3-3a.This suggested that Ca14-3-3a is most likely involved in the sucrose metabolism .Real-time PCR showed that the expression of Ca14-3-3a was continuously clined with the time in the Ralstonia solanacearum infected pep-per,it is inferred that it is also involved in the defense reaction of the plant .