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[目的]采用Taqman定量PCR技术检测转基因杂交大豆中外源nos终止子基因的拷贝数。[方法]以大豆凝集素基因为内参照基因,以非转基因大豆基因组DNA为内参照基因标准品,通过梯度稀释法分别求取了内参照基因和质粒DNA的Ct值与拷贝数对数值的相关性标准曲线方程,并通过将得到的Ct值代入标准曲线方程求取了样品的拷贝数。[结果]内参照基因标准曲线方程为y=-3.422x+35.201,R2=0.998;外源基因标准曲线方程为y=-3.348x+34.890,R2=0.999。nos终止子基因及其下游边界序列在转基因杂交大豆中为单拷贝。[结论]该研究为确定转基因大豆外源基因拷贝数提供了理论依据。
[Objective] To detect the copy number of foreign nos terminator gene in transgenic hybrid soybean by Taqman quantitative PCR. [Method] With soybean agglutinin gene as internal reference gene and non-transgenic soybean genomic DNA as internal reference standard, the correlation between Ct value and copy number logarithm of internal reference gene and plasmid DNA was obtained by gradient dilution method The standard curve equation was obtained and the copy number of the sample was obtained by substituting the obtained Ct value into a standard curve equation. [Result] The standard curve equation of internal reference gene was y = -3.422x + 35.201, R2 = 0.998. The equation of exogenous gene standard curve was y = -3.348x + 34.890, R2 = 0.999. The nos terminator gene and its downstream border sequence are single copies in the transgenic hybrid soybean. [Conclusion] The study provided the theoretical basis for determining the copy number of foreign gene in transgenic soybean.