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通过染色体步移方法从银杏(Ginkgo biloba L.)基因组中克隆到查尔酮合成酶基因(CHS)翻译起始位点上游1711bp的启动子序列。生物信息学分析表明,该启动子片段中存在多个顺式作用元件,包括紫外/蓝光响应单元、植物激素响应单元、真菌诱导元件、MYB结合位点、TATA-box和CAAT-box等。亚克隆了CHS转录起始位点上游1402bp序列,将其与GUS基因构建融合表达载体pBI121+CHSP,以pBI121-35S作为负对照,通过农杆菌(LBA4404)介导法分别转入烟草。结果表明,银杏CHS启动子序列能驱动GUS基因在烟草中的表达,表达具有组织特异性。GbCHSP的功能研究将有助于揭示银杏叶黄酮的积累与GbCHS基因表达的分子机理。
A 1711bp promoter sequence upstream of the chalcone synthase gene (CHS) translational start site was cloned from the genome of Ginkgo biloba L. by chromosome walking. Bioinformatics analysis showed that there are many cis-acting elements in the promoter fragment, including UV / blue response unit, plant hormone response unit, fungal induction element, MYB binding site, TATA-box and CAAT-box. The 1402bp sequence upstream of the CHS transcription start site was subcloned, and fused with GUS gene expression vector pBI121 + CHSP. PBI121-35S was used as a negative control and transformed into tobacco by Agrobacterium (LBA4404) mediated method. The results showed that GUS gene CHS promoter could drive the expression of GUS gene in tobacco and its expression was tissue-specific. The functional study of GbCHSP will help reveal the molecular mechanism of GbCHS gene expression and the accumulation of flavonoids in Ginkgo biloba.