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

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.

The quantitative determination of antiACC synthase gene RNA expression in transgenic Zixyphus jujuba plant is carradout by real time reversetranscription 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(R2=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 antiACC synthase gene is 4 50010 300 copies/g tissue.