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在已知GCYC基因部分序列基础上,通过改进的mTAIL-PCR方法克隆非洲紫罗兰Saintpauliaionantha两侧对称栽培种中CYC类基因的5′未知序列,并进而从两侧与辐射对称栽培种中分离得到苦苣苔科Gesneriaceae中第一组完整基因:SiCYC1A与SiCYC1B。对以上基因的核酸和氨基酸序列比较发现,SiCYC1A与SiCYC1B序列同源性很高,均含有完整的功能调控区域(即TCPdomain和Rdomain)并与模式植物金鱼草Antirrhinummajus中CYC基因同源。因此,这两个基因应具有正常功能,是功能上互补的冗余基因。令人意外的是在辐射对称花栽培品种中的这两个基因和两侧对称花栽培品种中对应基因的序列完全相同。经过对金鱼草以及相关类群辐射对称花突变体中CYC类基因序列的比较分析,推论在非洲紫罗兰中,SiCYC1A与SiCYC1B基因可能受上游未知的共同调控因子调控,该调控因子的改变是导致栽培品种中花对称性发生变化的主要原因。另外,对改进后的TAIL-PCR(mTAIL-PCR)的方法和过程进行了详细叙述,并对其技术特征和优势开展了简单的论述。
Based on the known sequence of GCYC gene, the 5 ’unknown sequence of CYC gene was cloned by modified mTAIL-PCR from symmetrical cultivars of Saintpaulia ionantha on both sides, and was isolated from the symmetrical cultivars The first complete gene of Gesneriaceae in the family Gesneriaceae: SiCYC1A and SiCYC1B. Comparison of the nucleotide and amino acid sequences of the above genes showed that SiCYC1A had high homology with the sequence of SiCYC1B and contained complete functional regulatory regions (ie, TCPdomain and Rdomain) and was homologous to the CYC gene in the model plant Antirrhinummajus. Therefore, these two genes should have normal functions and be functionally complementary redundant genes. What is surprising is that the sequences of the corresponding genes in these two genes and the bilaterally symmetrical flower cultivars in the radiation-symmetric flower cultivar are exactly the same. Based on the comparative analysis of the CYC gene sequences of the snapdragon and its related groups of radiant symmetrical flower mutants, it is concluded that the SiCYC1A and SiCYC1B genes may be regulated by the common upstream and downstream regulatory factors in the African violets, The main reason for the change of flower symmetry. In addition, the method and process of modified TAIL-PCR are described in detail, and its technical features and advantages are briefly discussed.