ECR1-like Gene Copy Number Variation Detection And SNP Analysis In Different Individuals Of Landrace

Objective:In order to investigate the molecular basis of the Erythrocyte complement receptor 1-like(ECR1-like)protein phenotype polymorphism,we studied single nucleotide polymorphism(SNP)and copy number variation(CNV)of different individuals ECR1-like gene in Landrace.In order to provide theoretical data for the study of ECR1-like structural polymorphisms in pigs and the molecular mechanisms that affect red blood cell immune adhesion function.Methods:In this study,41 Landrace as the subjects,the study of ECR1-like gene as target.The ECR1-like coding genes were analyzed by the bioinformatics analysis modules such as GENE,SNP,and BLAST in the NCBI database to predict the exon region and some SNPs of the ECR1-like gene.The SNP loci were genotyped by direct sequencing and allele-specific PCR(AS-PCR).Analysis of SNPs using Hardy-Weinberg equilibrium test and linkage disequilibrium test.Haploview 4.3 software was used to construct the haplotypes of ECR1-like gene SNPs.The ECR1-like gene copy number in each sample was detected by real-time fluorescence quantitative PCR technology and 2-??Ct relative quantitative analysis method,and the copy number difference was analyzed by using a single sample t-test in SPSS Statistics software.Results:(1)The analysis predicts that there are 23 exons in the ECR1-like gene and 62 SNPs are selected.After the prediction,the SNP(rs321524247)site located in exon 3 is a missense mutation,causing amino acid mutations.The AS-PCR genotyping method for this SNP was successfully established.(2)There were 7 missense SNPs in the ECR1-like gene detected by exon sequencing.The results showed that there were 6 missense SNPs in exon 5 and one missense SNP in exon 16.A total of 8 SNPs including the SNP(rs321524247)of exon 3 detected by AS-PCR were analyzed together,it was concluded that the two SNPs,rs321524247 and rs330380626,were in Hardy-Weinberg equilibrium and were completely linked(D'=1,r2=1),the haplotype AT was constructed as the major haplotype with a frequency as high as 0.915.(3)ECR1-like gene copy number range is 0.14-1.45,showing normal copy number and deletion of copy number.Among them,the ECR1-like gene in Landrace pigs was normal,accounting for 68.30%of the total,and the copy number was 31.70%of the total.The t-test analysis showed that the normal copy number and copy number deletion were significantly different(P<0.01).Conclusious:(1)There are more SNPs in ECR1-like exons,which indicates that ECR1-like is prone to mutations,leading to changes in protein structure,expression,and molecular weight.And there was a linkage disequilibrium between the detected SNPs.Multiple SNP mutations may occur causing a trait change.(2)ECR1-like has a loss of copy number.Theoretically,loss of ECR1-like copy number will result in a decrease in the protein expression,causes the body's immune system to decline.Therefore,prediction of ECR1-like copy number can be used as an indicator to detect pig erythrocyte immune level.