CAJ | 학술논문

用根据核苷酸结合位点(nucleotide binding site, NBS)和丝氨酸/苏氨酸蛋白质激酶域设计的2对简并性引物,以小麦-簇毛麦6VS/6AL易位系的cDNA为模板进行PCR扩增.扩增产物克隆到pGEM-T载体中,经测序,共获得具有NBS结构域特征的片段克隆9个和具有丝氨酸/苏氨酸蛋白质激酶域特征的片段的克隆1个.将克隆之间核苷酸序列同源性高于90%的克隆归为一类,把9个NBS片段分为6类.这6类抗病基因类似序列(resistance gene analogs, RGA)均具有阅读框,并与已克隆的小麦抗条锈病基因Yr10、大麦抗白粉病基因Mla1和Mla6、拟南芥的抗病基因RPS2以及其他一些抗病基因在NBS保守区内具有高度的同源性.用小麦中国春缺体-四体初步将它们分别定位于小麦第一、第二和第五部分同源群上.进一步用5′-RACE技术获得RGA N5的5′-端,发现其编码产物的N端还具有6个亮氨酸拉链(leucine zipper, LZ),与RPS2的N-端有较高同源性.
용근거핵감산결합위점(nucleotide binding site, NBS)화사안산/소안산단백질격매역설계적2대간병성인물,이소맥-족모맥6VS/6AL역위계적cDNA위모판진행PCR확증.확증산물극륭도pGEM-T재체중,경측서,공획득구유NBS결구역특정적편단극륭9개화구유사안산/소안산단백질격매역특정적편단적극륭1개.장극륭지간핵감산서렬동원성고우90%적극륭귀위일류,파9개NBS편단분위6류.저6류항병기인유사서렬(resistance gene analogs, RGA)균구유열독광,병여이극륭적소맥항조수병기인Yr10、대맥항백분병기인Mla1화Mla6、의남개적항병기인RPS2이급기타일사항병기인재NBS보수구내구유고도적동원성.용소맥중국춘결체-사체초보장타문분별정위우소맥제일、제이화제오부분동원군상.진일보용5′-RACE기술획득RGA N5적5′-단,발현기편마산물적N단환구유6개량안산랍련(leucine zipper, LZ),여RPS2적N-단유교고동원성.
Two pairs of degenerate primers were designed based on nucleotide-binding site (NBS) and serine/threonine kinase domain. PCR was performed with the primers and cDNA from the Triticum aestivum-Haynaldia villosa translocation line 6VS/6AL. Amplified products were cloned and sequenced. Nine clones with NBS and one with serine/threonine kinase domain were obtained. The NBS clones were classified to six groups according to their nucleotide sequence identities (90% or higher). These resistance gene analogs (RGAs) all have open reading frames (ORF), and their amino acid sequences show high similarity to Yr10 in wheat, Mla1 and Mla6 in barley, RPS2 in Arabidopsis and other resistance (R) genes with conserved motifs. They were preliminarily mapped on the chromosomes of homoeologous groups 1, 2 and 5 of common wheat by nulli-tetrasomic analysis. The 5′-end sequence of an RGA N5 was obtained by 5′-RACE PCR. It encodes six leucine zipper (LZ) and has high sequence similarity to RPS2.