CAJ | 학술논문

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.
종불결구백채CMS신충질중분리득도적일개cDNA-AFLP차이편단,채용RT-PCR화RACE기술성공극륭료일개α-미관기인적cDNA전장서렬,명명위TUBA2(DDBJ등록호위AB445012).서렬분석결과표명,해기인전장1 709 bp,최대개방열독광위1 353 bp,편마450개안기산서렬,여이공포적α-미관기인유교고적동원성.계통진화수분석발현,해기인재불동식물간구유고도보수성.Southern잡교표명TUBA2속우불결구백채다기인가족적일개단일극륭기인.실시정량RT-PCR검측표명,해기인재불육계중적표체량현저저우보지계,동시재불동조직화세포감수분렬불동시기해기인적표체량야존재명현차이.
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.