| 摘要: |
| [目的]研究烟草MARs序列对外源基因表达的影响,从中筛选出能明显提高目的基因表达水平的强MARs序列.[方法]用PCR方法从烟草基因组中克隆得到3个MARs序列(TM1、TM2和TM3),将这3个MARs序列分别顺向构建到植物表达载体pBI121 GUS基因表达盒两端,经农杆菌介导转化烟草,对获得的转基因植株体内的GUS酶活性进行了定量检测,分析MARs对GUS酶活性的影响.[结果](1)序列分析表明,TM2和TM3是两个新的MARs序列;(2)GUS酶活性的定量检测表明,TM1、TM2、TM3可以使GUS基因的平均表达活性分别提高1.4,5.1和1.3倍,但不同转化个体之间的表达水平有明显差异.[结论]虽然克隆到的3个MARs序列均可提高外源基因GUS的表达,但其并未降低转基因植株个体间外源基因表达水平的差异. |
| 关键词: 烟草 MARs序列 GUS基因 表达活性 |
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| 基金项目:泰安市科技局计划项目（2006D1076）；泰山医学院青年科学基金项目（2007QN57） |
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| Isolation and functional identification of tobacco MARs |
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| Abstract: |
| 【Objective】 The paper studied the effects of tobacco MARs on transgene expression and select strong MARs which can obviously increase transgene expression level. 【Method】 Three MARs （TM1，TM2 and TM3）were cloned from tobacco genome by PCR.Vectors pBI121 were modified with direct repeat MARs inserted on both sides of the reporter gene cassette and were transferred into tobaccos via Agrobacterium-mediated transformation method.GUS activity of the transgenic tobaccos was detected according to Jefferson and the effect of MARs on GUS activity was analyzed.【Result】 (1) Sequence analysis showed that TM2 and TM3 were two new MARs.(2)Quantitative detection of GUS activity for the transgenic tobaccos showed that TM1,TM2 and TM3 increased the average GUS activity by 1.4-folds,5.1-folds and 1.3-folds.But the expression differences among individual transformants were still obvious. 【Conclusion】 Three MARs could increase GUS gene expression level,but the expression differences among individual transformants were still obvious. |
| Key words: tobacco MARs GUS gene expression activity |
