Identification Of RyR Gene Family And Expression Analysis In Different Tissues Of Coilia Nasus

Ryanodine receptor（Ry R）belongs to a calcium channel protein,the main expression site is in the sarcoplasm and the endoplasmic reticulum membrane,and its main function is to store and release Ca²⁺from the cell during the excitation and contraction coupling process of myocardium and skeletal muscle.It is important ion in organisms,plays an important role in maintaining the metabolism.This ion can not only play a role in nerve regulation and the excitatory conduction of the muscle system,but also participate in important life activities such as the fertilization process,embryonic development,cell division,cell differentiation,and cell apoptosis.Coilia nasus is a migratory fish with economic value,and its genome information has been published,but there are few reports on the identification of gene family of transcription factors of Coilia nasus,but the research and analysis on the identification of Ry R gene family of Coilia nasus have not been reported.Adaptation to the changes of salinity and osmotic pressure is a necessary condition for successful migration,the Ca²⁺channels protein also make outstanding contributions in regulating osmotic pressure.Meanwhile,as one of the Ca²⁺channel proteins,we are research on the Ry R gene family and expression in different tissues of Coilia nasus.Firstly,we used Coilia nasus genome data for the study of material,using biological information to research the Ry R gene family of Coilia nasus.the physical and chemical properties,phylogeny,chromosome location,etc.of the obtained Ry R protein of Coilia nasus.Secondly,the full-length transcriptome of Coilia nasus was obtained by using three generation sequencing technology.In my paper,q PCR technology was used to analyze the tissue expression patterns of the three subtypes of Ry R gene in the gills,brain,ovary and testis of Coilia nasus,providing theoretical data for the research of the function of the Ry R gene in Coilia nasus.The main research results are as follows:1.According to the genomic data of Coilia nasus and the Ry R gene family sequences of zebrafish and other species,the Ry R gene family of Coilia nasus was searched in the genome of Coilia nasus.Eight Ry R genes were identified in the whole genome database of Coilia nasus.They were distributed on seven chromosomes of Coilia nasus,and there were two genes on chromosome 1.The size is about 550kd,and the PI value is between5.03 and 5.75.Of the 8 Ry R genes in Coilia nasus,Cn Ry R1、Cn Ry R3、Cn Ry R2 belong to the Ry R1,Cn Ry R4、Cn Ry R6、Cn Ry R8 belong to the Ry R2 and Cn Ry R5、Cn Ry R7 belong to the Ry R3.2.A total of 64.18Gb of clean data was obtained from the full-length transcript of Coilia nasus.739,989 cyclic consistent sequences（CCS）were obtained.There were 590,087full-length non-chimeric sequences（FLNC）.After clustering FLNC,168,407 consistent sequences were obtained.Then 165,413 high quality consistent sequences were obtained After redundancy removal,80,620 transcripts were obtained The non-redundant transcriptional integrity was evaluated,and there were 2,101 perfectly matched sequences.Sequence structure analysis of the newly discovered transcripts revealed 267,282 SSR sequences,92,178 complete ORF sequences and 1,418 Lnc RNAs,and a total of 1,393 Lnc RNA target genes were predicted.29,299 spliceable events;Functional annotation of 109,922 new transcripts was completed;7,421 transcription factors were predicted,mainly belonging to 20 gene families,of which ZF-C₂H₂,ZBTB,Homebox,BHLH and other gene families were the most.Full-length transcriptome data showed that 8 Ry R genes were identified at the gene level,which was consistent with the number identified.Twelve Ry R genes were found at the transcriptome level,including eight identified by genome and four other Ry R genes.3.The differential expression of Ry R gene in brain,gill,ovary and testis of Coilia nasus was studied.The expression of the three subtypes of the Ry R gene in the gills,brain,and gonads（ovary,testis）of Coilia nasus was detected by q PCR.The results showed that the Ry R1 gene was expressed in the gills and brain,but was not expressed in the ovary and testis;the expression of Ry R2 gene in the gill tissue of the seawater group was lower than that of the freshwater group,and the expression level of the brain tissue of the seawater group was higher.In the freshwater group,the expression levels of ovarian and testis tissues did not show any difference between the seawater group and the freshwater group;the expression level of Ry R3 gene in the gill,brain,and ovarian tissues in the seawater group was higher than that in the freshwater group,and the gill tissue showed extremely significant difference（P<0.01）,showing significant difference in brain and ovarian tissues（P<0.05）.