Application of cl5547.contig2 gene as an internal reference gene in real-time fluorescent quantitative PCR analysis of pumpkin gene expression

Abstract

The invention belongs to the technical field of gene expression analysis, and concretely relates to application of CL5547.Contig2 gene to pumpkin gene expression real-time fluorogenic quantitative PCR analysis as a reference gene. The gene expression level and stability are compared at whole genome level by utilizing pumpkin transcriptome data, 10 genes with stable expression are selected, also 2 commonly-used conventional reference genes in pumpkin are selected as contrast, and the expression stability of the 12 genes in different tissue organs of two pumpkin varieties under conditions of abiotic stress, biotic stress and plant-growth regulator processing (GA3, ABA, NAA and ETH), and CAT1 gene and CAT2 gene are used to perform verification, so that the CL5547.Contig2 gene is confirmed to have the highest expression stability, and is an optimum reference gene for pumpkin gene expression real-time fluorogenic quantitative PCR analysis.

Description

technical field [0001] The invention belongs to the technical field of plant molecular biology, and in particular relates to an internal reference gene required in the pumpkin gene expression analysis process. Background technique [0002] Gene expression analysis is an important means to study gene function and transcriptional regulation. Real-time fluorescent quantitative PCR (qRT-PCR) is widely used in the quantitative analysis of gene expression because of its high sensitivity, high specificity, high accuracy and low cost. In qRT-PCR analysis, in order to obtain the real difference in the expression level of the target gene, it is necessary to eliminate the differences in the RNA yield, quality and reverse transcription efficiency of different samples. To eliminate the difference between samples, the internal reference gene is usually used for correction and standardization, that is, by measuring the expression of the target gene and the internal reference gene respecti...

AI Technical Summary

Problems solved by technology

At present, traditional internal reference genes are mostly used in pumpkin gene expression analysis. These traditional internal reference genes have not been systematically verified and have poor stability, resulting in poor accuracy of pumpkin gene expression analysis.

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