【摘要】 从不结球白菜CMS新种质中分离得到的一个cDNA-AFLP差异片段,采用RT-PCR和RACE技术成功克隆了一个α-微管基因的cDNA全长序列,命名为TUBA2(DDBJ登录号为AB445012)。序列分析结果表明,该基因全长1 709 bp,最大开放阅读框为1 353 bp,编码450个氨基酸序列,与已公布的α-微管基因有较高的同源性。系统进化树分析发现,该基因在不同植物间具有高度保守性。Southern杂交表明TUBA2属于不结球白菜多基因家族的一个单一克隆基因。实时定量RT-PCR检测表明,该基因在不育系中的表达量显著低于保持系,同时在不同组织和细胞减数分裂不同时期该基因的表达量也存在明显差异。更多还原

【Abstract】 On the basis of one cDNA-AFLP differential expression fragment isolated from the new germplasm of non-heading Chinese cabbage(Brassica campestris ssp.chinensis Makino).The full length cDNA of the gene were cloned by reverse transcription polymerase chain reaction(RT-PCR) and rapid amplification of cDNA ends(RACE).The gene named TUBA2(DDBJ accession No.AB302891),the length of its cDNA was 1 709 bp,which include an open reading frame(ORF) with 1 353 bp encoding 450 bp amino acid residues,and showed high degree of homology to other plants α-tubulin proteins.Phylogenetic analysis revealed the evolutionary conservation of this protein among different plants.The Southern blot result suggested that the cloned and characterized gene TUBA2 was a single copy gene and belonged to a multiple gene family(about two to four α-tubulin genes) in non-heading Chinese cabbage.Real-time quantitative PCR analysis revealed the gene TUBA2 was expressed in a significantly lower level in CMS line than that in the maintainer and there were significant differences in gene expressions at different organs and all of the stages of the microsporogenesis.更多还原