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本研究主要探讨ipt基因对矮牵牛遗传转化不定芽诱导影响及拟南芥热激启动子hsp18.2驱动重组酶基因flp的热诱导外源基因删除表达载体在矮牵牛中的基因删除效果。本研究中将植物表达载体pBin-hsp18.2:flp-35S:ipt及对照载体pBin-hsp18.2:flp遗传转化矮牵牛,以获得转基因植株,分析比较不定芽的诱导和转基因植株进行的热激基因删除。研究结果表明,ipt基因可促进矮牵牛遗传转化过程中不定芽的诱导,其不定芽诱导率为21.5%,显著高于对照的8.7%。在44℃,6h,热激6次的条件下,转基因矮牵牛植株表型恢复正常,经GUS蛋白表达分析及PCR、RT-PCR检测,证明外源基因已经被删除。转基因矮牵牛基因删除效率最高可达43.8%。本研究为ipt基因在一些遗传转化困难植物转基因中的应用奠定了基础。
Abstract：In this research,we studied the influence of petunia shoot regeneration when transformed with ipt gene,and gene deletion effect of the heat shock inducible "gene deletion" construction in which flp gene was driven by the promoter hsp18.2.We transformed constructions of pBin-hsp18.2:flp-35S:ipt and pBin-hsp18.2:flp into petunia axilaris,to gain transgenic plant.We analyzed shoot regeneration between pBin-hsp18.2:flp-35S:ipt and pBin-hsp18.2:flp transformed plant,studied gene deletion efficiency with heat shock.The results showed that ipt gene can enhance adventitious shoots regeneration of petunia.The shoots inductivity of pBin-hsp18.2:flp-35S:ipt lines is 21.5% evidently higher than pBin-hsp18.2:flp lines which are 8.7%.Furthermore,the phenotype of transgenic plant became recovery by heat shock,which treatment conditions was heat shock under temperature of 44℃ for 6 h,and heat shock for 6 times.It proved the exogenous gene has been deleted in transgenic plants by GUS and PCR analysis.The highest gene deletion efficiency is 43.8% in this reasearch.This study provide evidence for application of ipt gene in transformation with plants that difficult to transform.
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基本信息:
中图分类号:S681.9
引用信息:
[1]李岩,赵德刚.ipt基因促进矮牵牛遗传转化效率及热激启动子驱动基因删除[J].基因组学与应用生物学,2011,30(02):145-151.
基金信息:
科技部国际科技合作项目(2007DFA31260);科技部国家科技支撑计划项目(2007BAD59B06);科技部国家科技支撑计划项目(2008ZX08010-003)共同资助
2011-04-28
2011-04-28