Correlation Between NLRP3 Gene Non-coding Region Polymorphism And Susceptibility To Mycoplasma Pneumonia In Sheep

Objective:(1)The prevalence of Mycoplasma pneumonia in Kazakh sheep and Suffolk sheep in Ili area of Xinjiang was identified through the clinical manifestations and pathological changes of Mycoplasma ovine pneumoniae infection,combined with serological detection methods,and the number of Mycoplasma pneumoniae in sheep populations of different breeds was calculated.infection rate.(2)The SNP loci in the non-coding region of the NLRP3 gene were screened by PCR detection method,and the genotypes of different SNP loci were analyzed to explore the genetic polymorphism of NLRP3 inflammasome-related genes in different sheep breeds.Correlations of susceptibility to Mycoplasma ovine pneumonia.Methods:(1)In this study,a total of 800 serum samples of two sheep breeds(Kazakh sheep and Suffolk sheep)were collected from some large-scale sheep farms in Yili,Xinjiang,and the Mycoplasma ovine pneumonia enzyme-linked immunosorbent assay(ELISA)detection kit was used.M.ovine pneumonia antigen and antibody serological testing was performed.(2)On this basis,the direct sequencing method was used to detect the single nucleotide variation in the non-coding region of the candidate gene NLRP3 gene,and the genetic diversity of Kazakh sheep and Suffolk sheep was compared according to the sequence comparison results of DNAMAN software and Chromas software.The genotype distribution was summarized,and the correlation analysis of the infectivity of Mycoplasma ovine pneumonia was carried out on the relevant mutation sites.Result:(1)The serological investigation of Mycoplasma ovine pneumonia in some large-scale sheep farms in Ili area of Xinjiang found that there were different degrees of Mycoplasma ovine pneumonia(MO)infection among different sheep breeds in Ili area of Xinjiang.The positive rate of MO-Ab in some largescale sheep farms in Yili area was 14.88%,the positive rate of MO-Ag was 21.00%,of which the positive rate of MO-Ab in Kazakh sheep was 26.00%,and the positive rate of MO-Ag was 22.91%;MO-Ab positive rate was 7.34%,MO-Ag positive rate was 19.71%.(2)Analyze the SNP sites in the non-coding region of the NLRP3 gene,and screen out C38233659 T,C38233866T,T38233881 C,A38188131C,G38188003 A,T38188153C,A38188237 C,a total of 7 non-coding region SNP sites,of which the base C38233659 T of the 3’UTR was detected.Mutation C→T,three genotypes of CC,CT and TT were detected in both sheep breeds.Among them,the dominant genotype of Suffolk sheep was CC,and the dominant allele was C;also in the C38233866 T site of 3′UTR The base mutation C→T was detected,and three genotypes of CC,CT and TT were detected in both sheep breeds.The dominant genotypes of Kazakh sheep and Suffolk sheep were both CC,and the dominant alleles were C;but The T38233881 C locus of the3′UTR was found only in the Kazakh sheep population with the base mutation T→C,the dominant genotype was TT,and the dominant allele was T.A base mutation A→C was detected at the A38188131 C site of the 5′UTR,and three genotypes of AA,AC and CC were detected in both sheep breeds.The dominant genotypes of Kazakh sheep and Suffolk sheep were both AA.All alleles are A;base mutation G→A was detected at the G38188003 A site of 5′UTR,and three genotypes of GG,GA and AA were detected in both sheep breeds.The advantages of Kazakh sheep and Suffolk sheep The genotypes were all GG,and the dominant alleles were all G.In addition,the base mutation T→C was detected at the T38188153 C site of the 5′UTR,the dominant genotype was TT,and the dominant allele was T;the base mutation A→C was detected at the A38188237 C site of the 5′UTR,and the dominant gene was The type is AA,and the dominant allele is A.The mutation of the above two loci only appears in the Suffolk sheep population,but not in the Kazakh sheep population.There was a significant difference between the genotype frequencies of MO-Ag positive and MO-Ag negative in Kazakh sheep at the A38188131 C site of the 5′UTR of NLRP3 gene(P<0.05).Conclusion: The infection of Mycoplasma ovine pneumonia exists among different sheep breeds in some large-scale sheep farms in Yili,Xinjiang,and the infectivity of Kazakh sheep is higher than that of Suffolk sheep.The test results show that the infectivity of local breeds is higher than that of imported breeds.The A38188131 C locus of the 5′UTR of NLRP3 gene may be related to the susceptibility of Kazakh sheep to MPS,so this locus can be used as a molecular genetic marker locus for the susceptibility of M.ovine pneumonia,which is the breeding of new lines against M.ovine pneumonia.Provide theoretical support.