Internal reference gene act/gapdh of sisal and its application

Abstract

The present invention provides sisal ACT and GAPDH gene sequences, which can be used as sisal internal reference genes, and simultaneously provide sisal ACT and GAPDH as specific primers for internal reference genes, and establish a qRT-PCR method based on SYBR Green fluorescent dye technology, for Subsequent use of real-time fluorescent quantitative PCR provides stable and reliable internal reference genes and technical methods for gene expression analysis under stress conditions such as Phytophthora sisalum infection, low temperature stress, salt stress, and wound treatment.

Description

technical field [0001] According to an existing nucleic acid sequence, the present invention screens out an internal reference gene stably expressed in sisal under adverse conditions such as Phytophthora nicotiana infection, low temperature stress, salt stress and wound treatment, and belongs to the technical field of plant molecular biology. Background technique [0002] Sisal is one of the unique economic crops in the tropics of our country. Sisal is not only the main tropical fiber raw material, the sisal stem core is the main raw material for brewing tequila, and the sisal hemp residue contains high saponin, which can be used for pharmaceuticals , sisal is also an important biomass energy, and in recent years, as a model plant for sedum metabolism, sisal has very important theoretical and economic value. [0003] With the rapid development of sequencing technology and bioinformatics, the use of omics to study gene expression profiles under a certain stress has become a r...

AI Technical Summary

Problems solved by technology

[0003] With the rapid development of sequencing technology and bioinformatics, the use of omics to study gene expression profiles under a certain stress has become a routine method for molecular biology research, but omics is only a preliminary analysis of gene expression under a certain stress However, the results need to be verified by fluorescent quantitative RT-PCR; in addition, the spatio-temporal expression of genes has become one of the main contents of gene function research, and the above studies require relatively stable expressed genes as internal reference genes; With the in-depth development of hemp molecular biology, some functional genes have been successfully isolated. At present, there are many nucleic acid sequences of agave hemp in NCBI public data, but there are few internal reference gene sequences, only actin1 (232bp) and 18S rRNA (186bp) Because the expression level of internal reference genes may change under different conditions, different tissues, different periods and different varieties, this will lead to unstable research results and reduce the reliability of the results. Therefore, the screening of sisal is different. Stable expression of internal reference genes under adverse conditions is the premise of sisal gene function research; this not only improves the current situation of few internal reference genes in sisal, but also provides a guarantee for the reliability of the research results of sisal gene expression levels

Images