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由ACC合成酶反义基因转化所得枣树其RNA表达量的测定
Quantitation of Anti-ACC Synthase Gene RNA Expression in Transgenic Zixyphus jujuba by Real- time Reverse-transcription PCR Assay
【摘要】 利用real timeRT PCR对转基因枣树中的anti ACSG的RNA表达量进行了定量测定.样品扩增产物荧光量随着扩增循环数的增加而呈现"S"形增加;经熔解曲线分析,其扩增产物中未检出非特异性产物和引物二聚体;扩增产物的CT值和模板cDNA起始浓度两者之间呈线性相关(R2=0.9929);转化体中anti ACSGRNA表达量约为4500~10300拷贝/g组织.
【Abstract】 The quantitative determination of antiACC synthase gene RNA expression in transgenic Zixyphus jujuba plant is carradout by real time reversetranscription PCR. Results reveal that the amplification products fluorescence values of samples in crease in the form of"S"with the increase of cycle numbers. A melting curve analysis shows that, the quantity of unspecific PCR products have witch fewer primer dimers than other amplification products. Results show that there exists a linear correlation(R2=0.992 9) between the threshold cycle values of amplification products and the starting concentration of template cDNA. In the transformants, the expression quantitation of antiACC synthase gene is 4 50010 300 copies/g tissue.
【Key words】 real-time PCR; RT-PCR; anti-ACC synthase gene RNA; quantitative assay; Zixyphus jujuba;
- 【文献出处】 湖南农业大学学报(自然科学版) ,Journal of Hunan Agricultural University , 编辑部邮箱 ,2003年02期
- 【分类号】Q943.2
- 【被引频次】19
- 【下载频次】178