摘要
建立了番茄子叶高频再生体系,优化了农杆菌介导的外源基因转化番茄的条件,将IPT和etr1 1双基因导入番茄中,以除草剂PPT作为筛选标记,得到3株转化再生植株.通过PCR,RT PCR检测证明etr1 1和IPT基因已整合到转基因番茄植株的基因组中,并在转录水平有一定表达.
The high-frequency regeneration system of Cherry tomato (Lycopersicum esclentum Mill) was established.Taking orthogonal experiment, the optimal combination and concentration of hormones were BA 1.0?mg/L and IAA 0.2?mg/L.The agrobacterium-mediated transformation system was optimized: OD_(600)0.1～0.2 was the fitful bacteria concentration for invading; the explants and the bacteria had been best co-cultured for 48 hours; choosing after being cultured in medium without PPT for 7 days were better; the optimal choice press is PPT 3?mg/L.Both IPT and etr1-1 genes were transferred into tomatoes with agrobacterium-mediated transformation.With PPT resistance as choice mark, three transgenic plants were regenerated with success.The transgenic tomato plants were confirmed by PCR analysis and RT-PCR analysis.By PCR analysis,the results indicated that the foreign genes had been integrated into the genome of cherry tomato.The RT-PCR analysis showed that the etr1-1 gene had been transcripted.
出处
《南京师大学报（自然科学版）》
CAS
CSCD
2004年第3期83-87,共5页
Journal of Nanjing Normal University(Natural Science Edition)
