Study On TPM2 And TPM3 Gene Expression Patterns And Functions Of Bactrian Camel

As an ancient and unique animal,Bactrian camel has inestimable research value.Bactrian camel belongs to the typical spermatozoa-induced platoon animal,with obvious seasonal estrus characteristics.The different living habits of the production season and the non-production season make the reproductive physiology of the Bactrian camel different.Based on the use of proteomics technology in the early stage of this research team,it was found that muscle growth and development related differences between tropomyosin 2(TPM2)and tropomyosin 3(TPM3).The structure and function of tropomyosin 2 and tropomyosin 3 are both similar and specific.While encoding muscle isomers such as smooth muscle and skeletal muscle,it also produces different effects on cells by encoding muscle fibroblast isomers.In order to further reveal the role of TPM2 and TPM3 genes in Bactrian camel muscles at different production periods.In this study,the Bactrian camel in the production season was used as the research object.The TPM2 and TPM3 genes were cloned and sequenced by RT-PCR technology to analyze the basic functions of the TPM2 and TPM3 genes.To verify the specific expression of TPM2 and TPM3 in the production season of Bactrian camel muscles by immunofluorescence,immunohistochemistry,qRT-PCR,and Western blot,and explore the tissues of TPM2 and TPM3 in the production and nonproduction season expression pattern.Based on camel muscle fiber cells,the effect of interfering sequences on the apoptosis of muscle fiber cells was explored by transfecting siRNA.Result:1.The full-length CDS of the Bactrian camel TPM2 gene is 963 bp,and the full-length CDS of the TPM3 gene is 521 bp.They are acidic proteins.2.The mRNA and protein of TPM2 gene and TPM3 gene were expressed in the muscle tissues(smooth muscle,skeletal muscle,cardiac)of Bactrian camel during non-reproduction season and reproduction season,and the expression was up-regulated in Bactrian camel muscle during reproduction season(P<0.01).The expression in skeletal muscle is significantly higher than other tissues(P<0.01).3.The expression levels of TPM2 gene mRNA and protein in the experimental group transfected with siRNA sequences were significantly lower than those in the negative control group,and the expression level was the lowest at 24 hours after transfection(P<0.01).The expression level of Bcl-2 gene and TPM2 gene had a significant decrease trend at 24 h and 48h(P<0.01),while the expression level of Bax gene increased significantly at 24 h,48h(P<0.01)4.The expression levels of TPM3 gene mRNA and protein in the experimental grouptransfected with siRNA sequences were significantly lower than those in the negative control group,and the expression level was lowest at 48 hours after transfection(P<0.01).The expression level of Bcl-2 gene decreased significantly at 24 h,48h(P<0.01),and the expression level of Bax gene increased significantly from 24 h,48h(P<0.01)In summary,the TPM2 gene and TPM3 gene have different expressions in different muscle tissues of Bactrian camel.They participate in the muscle contraction process and have a certain effect on the apoptosis of muscle fibroblast.The mechanism of growth and development provides a theoretical basis.