拟南芥逆境诱导型启动子rd29A的克隆及活性检测

Cloning of stress-inducible promoter rd29A from Arabidopsis thaliana and its activity detection in transgenic tobacco

以拟南芥基因组DNA为模板,通过特异PCR扩增,克隆逆境诱导表达启动子rd29A。序列分析表明,该启动子与NCBI上已报道rd29A的启动子有99·64%的同源性,其序列含有干旱诱导表达元件DRE和ABA作用元件ABRE等顺式作用元件。用rd29A启动子替换pBI121载体上的35S启动子构建新的载体pBrd-GUS,并以农杆菌介导法将其转入烟草。转基因烟草的GUS组织化学染色及PCR分析结果表明,在低温、干旱及高盐等胁迫诱导下,rd29A启动子可增强GUS基因表达,因此,rd29A启动子可应用于植物抗逆基因工程研究。

Stress inducible promoter rd29A was cloned by PCR from Arabidopsis thaliana genomic DNA. Sequence analysis showed that this promoter was 99.64 % homologous with reported promoter rd29A in NCBI and contained several sis-acting elements including dehydration responsive element （DRE） and ABA responsive element （ABRE）. Promoter 35S of vector pBI121 was replaced by promoter rd29A, thus vector pBrd - GUS was constructed and transferred to tobacco by Agrobacterium tumefaciens system. PCR and GUS histochemical stain of transgenic tobacco showed that promoter rd29A induced by low temperature, dehydration and high salt stress strengthened expression of gene GUS. Therefore, promoter rd29A could be used to plant gene engineering for stress resistance improvement.