Double fluorescene report gene detection experiment
Experimental meaning
1. It can be widely referenced by target gene detection of miRNA, and it can be judged whether or not miRNA is combined with target genes, or it can quantify the inhibitory effect of miRNA to the purpose of interest.
2 Transcription factor ( transformer factor) There is a DNA binding domain that can be covalently bonded to a specific sequence (cis-acting element) in the target promoter, thereby regulating The expression of genes, the two-fluorescene reporting system is an important means of detecting such transcription factors and gene specific sequences, and is a common method of eukaryotic gene expression regulation.
Experimental principles and processes Dibluozyme refers to the lifelic luciferase and sea luciferase, firefly luciferase yellow green, wavelength 550-570 nm, sea kidney fluorescein The enzyme is blue, and the wavelength is 480 nm. The reporter gene experiment is to insert the specific fragment of the target latch into the front side of the fluorescent enzyme expression sequence, and construct a double reporter gene plasmid with a target promoter fragment, and then the transcription factor to the transcription factor to the plasmid and report genotyl plasmids HEK293 Cellular or other related cell lines, if the transcription factor can activate the promoter, report gene expression, and the intensity of the fluorescerase is proportional to the intensity of the transcription factor. The luciferase substrate is added to detect the interaction between the transcription factor and the target starter according to the fluorescence intensity. The luciferase reporter gene can be constructed in the same grain or construct is two plasmids, ie transcriptional factor plasmids and target starter fragment plasmids.
Experiment cycle: 4-6 weeks
Delivery standard: Double fluorescene report genetic testing, identification results report, analysis report, experimental report (reagent, Consumables, Experimental Step)