Exploited Of Immune-related Gene And Constructed Of Microarray In The Commom Carp(Cyprinus Carpio)

With the increasing density of aquaculture, the invasion of the disease has seriouslyhampered the aquaculture health development, thereby increasing the diseaseresistance of the fish on the large-scale sudden illness to become the focus of thestudy was notcomment EST sequences258,174,444carp immune-related sequenceswere identified gene function GO annotations, known nucleic acid sequence48thecarp immune function gene total of the carp immune function sequence492,492genes selected300gene primer design, amplification conditions screening to return atotal of264common carp immune-related functional gene primers.This experiment when the above-mentioned264pairs of primers for PCRamplification product as a probe, probe concentration of the gene chip0.5μg/μLPreferably, the point-like effect; spotting humidity of55points200M diameter of thedot pitch300μm point rounded, uniform size, and distributed evenly without cross;the test sample OD260/OD280value range of1.8to2.0, concentration300-500ng/μlwhen the hybridization the best; hybridization sample and hybridization solutionmixing ratio of1:6, hybridization temperature of55°C when the hybrid the besthybrid background is weak; differences in gene expression using the prepared genechip not spleen tissue the disease mirror carp and infected mirror carp detectionapplication QuantArray of microarray analysis software to analyze two the differencebetween the starting amount of sample RNA ratio analysis, identified by thedifferential expression of genes differentially expressed genes found in the two signalpaths Dendritic&Antigen Presenting Cell and antiviral response. This study throughon carp immune-related function gene of the Notes and Get research carp immunemechanism provides a theoretical basis, and carry out the construction of the genechip filter out264pairs of primers material, to optimize the preparation conditions forthe use of gene chips this high-flux fast means of detection of gene expressionpremise, not infected mirror carp and infected mirror carp out of the use ofimmune-related functional gene chip differential expression detection, proving thefeasibility of immune function gene chip application in the study of carp immunemechanism.