| Studies have shown that many factors can affect the growth and development of skeletal muscle,this study aims to solve the problems of low meat production rate,small body size and slow growth of cattle in China.The meat production performance of livestock and poultry is determined by muscle growth and development,from the embryonic stage to adulthood will be regulated by multiple genes,the research group in the early stage of the use of copy number variation,the gene screening,select the genes related to muscle growth and development.Based on this,this experiment took DYNC1I2 as the target gene,by interfering with DYNC1I2 and constructing an overexpression vector,and using q PCR,Westen Blot,CCK-8,Ed U,immunofluorescence and other techniques to explore its regulatory role in the proliferation and differentiation of bovine myocytes.In addition,real-time fluorescence quantification and SPSS correlation analysis were used to detect the copy number variation of DYNC1I2 gene in five cattle populations,and to explore the correlation between growth traits and copy number variation of beef cattle.The main research results are as follows:1.The distribution of DYNC1I2-CNV in cattle population and the correlation analysis with growth shapeThis study collected blood samples from 5 Chinese cattle breeds(Xianan cattle,Qinchuan cattle,Pinan cattle,Yunling cattle and Guyuan cattle),a total of 600 samples.Except for Guyuan cattle,the other4 cattle had one or more blood samples.Individual growth traits were significantly associated with copy number variation.In duplication individuals,this locus was significantly correlated with the body length of Pinan cattle(P < 0.05)and with the breast depth of Yunling cattle(P < 0.05);in deletion individuals,this locus was significantly associated with Xianan cattle The cross height of cattle was significantly correlated(P<0.05),and it was significantly correlated with the chest width,waist angle width and ischial end width of Qinchuan cattle(P<0.05).2.DYNC1I2 gene promotes the proliferation and differentiation of bovine myocytesThe test found that the gene was expressed in five tissues of heart,liver,lung,kidney and muscle during the fetal cattle period and the adult cattle period,indicating that the gene may play a wide role in the body.The expression regularity results of the two periods of 24-month-old bovine period and24-month-old adult bovine period showed that the expression level of fetal bovine period was higher than that of adult bovine period in muscle tissue.In terms of proliferation,after artificial interference and overexpression of DYNC1I2 gene,q PCR and Western Blot results showed that the expression levels of PCNA,CDK2 and Cyclin E1 were significantly inhibited after interference with this gene,and overexpression could significantly increase;The results of CCK-8 and Ed U showed that after interfering the gene,the cell activity decreased and the proliferation number of the cells decreased,and the result of overexpression was the opposite of the interference result.In terms of differentiation,interfering with this gene can interfere with the expression of Myo G gene,and overexpression can enhance the expression levels of Myo D and Myo G.In conclusion,DYNC1I2 gene can promote cell proliferation and differentiation.To sum up,It is clarified that DYNC1I2-CNV significantly affects some growth traits of cattle,and it is revealed that DYNC1I2 gene affects the proliferation and differentiation of cattle muscle cells.This study provides a new target gene for the growth and development of cattle,provides theoretical materials for exploring the genetic mechanism,and provides important information for beef cattle breeding.candidate molecular markers. |
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