普通小麦 TaPAPA1基因克隆与特异性标记开发

摘要

研究发现锌指（Zinc－finger HIT，Zf－HIT）蛋白在植物的抗逆性方面有重要的调控作用。本研究从普通小麦中克隆到一个含有 PAPA －1－like[Pim －1－associated protein －1（PAP －1）－associated protein －1－like]和 Zf －HIT 结构域的蛋白质基因，命名为 TaPAPA1。通过测序和序列分析，得到三条 gDNA 和三条cDNA 序列，均包含6个外显子和5个内含子。根据序列间的差异位点设计基因组特异引物，用中国春小麦缺体—四体系将该基因定位在小麦第五同源群。TaPAPA1－5A、TaPAPA1－5B 和 TaPAPA1－5D 的编码序列长度分别为1473、1473 bp 和1470 bp，分别编码490、490和489个氨基酸。在5B 染色体检测到该基因的一个 SNP 位点，并开发 AS －PCR 标记 TaPAPA1－5B －AS1／2。利用 RIL（Recombinant inbred line）群体，把该标记定位在 D －1207571和 D －1100783之间，遗传距离分别是0．51 cM和0．71 cM。%Zinc -finger HIT protein plays an important role in the regulation of plant defense and stress response.In this study,a gene named TaPAPA1,who encoded one protein containing PAPA -1 -like [Pim -1 -associated protein -1 (PAP -1 )-associated protein -1 -like]and Zf -HIT (Zinc -finger HIT)domain, was cloned from wheat.Through sequence analysis,three cDNA and three gDNA sequences were got,which all had six exons and five introns.According to different sites among three gDNA sequences,three pairs of genome special primers were designed,and these genes were located in homologous group 5 using nullisomic -tetrasomic lines of Chinese Spring.The coding sequences of TaPAPA1 -5A,TaPAPA1 -5B and TaPAPA1 -5D were 1 473,1 473 bp and 1 470 bp,which encoded polypepetides with 490,490 and 489 amino acids,respectively. According to the SNP locus in TaPAPA1 -5B,the AS -PCR marker TaPAPA1 -5B -AS1 /2 was developed.U-sing a RIL (recombinant inbred line)population,the marker was located between marker D -1207571 and D -1100783 on 5B by linkage analysis with the genetic distance of 0.51 cMand 0.71 cM,respectively.