半定量RT-PCR方法检测NaCl胁迫下杂花苜蓿mvNHX基因表达及体系优化

Expression of mvNHXgene in Medicago.varia by Semi-Quantitative RT-PCR Systems under NaCl Stress and Its System Optimization

【目的】优化半定量RT-PCR反应体系,并用优化体系检测杂花苜蓿mvNHX基因的表达情况。【方法】提取用150 mmol/L盐浓度、处理不同时间的苜蓿叶片组织总RNA;反转录得到cDNA;利用优化半定量RT-PCR法检测。【结果】检测到mvNHX基因随胁迫时间基因表达量总体上逐渐升高。【结论】通过半定量RT-PCR法检测基因在胁迫条件的表达量的变化。同时对该方法进行优化,为进一步研究该基因克隆及其功能验证奠定了一定的实验基础。

【Objective and Method】In this experiment,expression status of Medicago.varia mvNHX gene in 150 mmol/L NaCl for different times were detected under the stress treatment and appropriate semi-quantitative RT-PCR system was established.The total RNA of leaves tissue were extracted by 150 mmol/L NaCl at different time,cDNA were reversely transcripted and assayed by semi-quantitative RT-PCR.【Result】Detected mvNHX gene expression levels increased gradually in general with the stress time.【Conclusion】The amount change for gene expression were detected by semi-quantitative RT-PCR method under stress conditions.At the same time,the method was optimized to further study gene cloning to provide experiment basis for its functional verification.