多聚ADP核糖聚合酶(PARP)受基因毒剂的特异性诱导。将拟南芥(Arabidopsis thaliana)AtPARP1基因上游长2 179 bp的启动子片段插入到质粒pAKK687的β-葡萄糖醛酸糖苷酶(GUS)报告基因上游, 转化拟南芥。GUS组织化学染色结果表明, GUS报告基因仅在苗龄3–5天的拟南芥根部及花发育早期的雄蕊中表达; 1.5 μg·mL–1博莱霉素与22 μg·mL–1丝裂霉素联用强烈诱导了GUS报告基因的表达(尤其在拟南芥的幼苗和果荚中)。进一步降低抗生素浓度, 发现单独使用1 μg·mL–1博莱霉素对GUS报告基因也具较强的诱导活性, 且对拟南芥幼苗的生长无影响。上述结果表明, AtPARP1启动子是一个新型的具较大应用潜力的抗生素诱导型启动子。
Poly ADP-ribose polymerase (PARP) is induced specifically by genotoxin. We cloned the 2 179 bp promoter sequence of Arabidopsis AtPARP1 gene into a pAKK687 vector to drive the expression of a β-glucuronidase (GUS) reporter gene for Arabidopsis transformation. GUS staining revealed the GUS reporter gene expressed only in the basal root of 3 to 5-day-old seedlings and in the anther at early flowering stage. An amount of 1.5 μg·mL–1 bleomycin combined
with 22 μg·mL–1 mitomycin could strongly induce the expression of the GUS reporter gene, especially in young seedlings and young siliques of Arabidopsis. An amount of 1 μg·mL–1 bleomycin was enough to induce the promoter and had no adverse effects on Arabidopsis seedling growth. The AtPARP1 promoter may be a new antibiotic-inducible promoter with application potential.
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