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为了预测抽薹相关基因BrcuDFR-like/BrcuAXS的功能,通过PCR和RACE的方法克隆了菜薹BrcuDFR-like/BrcuAXS基因的cDNA和gDNA全长序列。结果表明:该基因编码区全长1332bp,编码312个氨基酸残基。对应的gDNA全长为2460bp,含有8个外显子和7个内含子,内含子总长为1042bp,其中第3个内含子最长,为401bp。内含子中含有多个基本转录元件和顺式作用元件,如光应答元件、赤霉素响应元件、参与抗性和胁迫应答元件、热响应元件、WRKY转录因子的结合位点及干旱胁迫元件MYB转录因子结合位点等。利用半定量RT-PCR分析表达模式,发现BrcuDFR-like/BrcuAXS随菜薹花芽形态逐步建成直至抽薹开花,其表达量逐渐增强,与其它物种DFR-like基因的表达模式更吻合,由此预测该基因在菜薹生长发育阶段编码DFR-like酶的可能性大于编码AXS的可能性,其功能可能与菜薹营养分生组织向花分生组织转变有关。
In order to predict the function of bolting-related gene BrcuDFR-like / BrcuAXS, the cDNA and gDNA full-length sequences of BrcuDFR-like / BrcuAXS gene were cloned by PCR and RACE. The results showed that the coding region of this gene was 1332bp in length and encoded 312 amino acid residues. The corresponding full-length gDNA was 2460bp, containing 8 exons and 7 introns. The total length of introns was 1042bp, of which the longest intron 3 was 401bp. Intron contains a number of basic transcriptional elements and cis-acting elements such as light-responsive elements, gibberellin responsive elements, resistance and stress response elements, heat-responsive elements, WRKY transcription factor binding sites and drought stress element MYB Transcription factor binding site and so on. The expression pattern of BrcuDFR-like / BrcuAXS was gradually established with the flower bud morphology of Cauliflower by semi-quantitative RT-PCR. The expression level of BrcuDFR-like / BrcuAXS was more consistent with the expression pattern of DFR-like genes in other species. The gene is more likely to encode DFR-like enzymes than cabbage in its growth and development stage, and its function may be related to the transformation of vegetative meristems into flower meristems.