Analysis Of MRNA Expression Profile In Mammary Epithelial Cells Of Sheep With Four-nipples By Over-expressed DEPDC7 Gene

DEPDC7 has two potential functional domains,the DEP domain and Rho GAP domain,containing BRCC 3-like proteins,which play important roles in signal transduction,cell polarity determination,and cell anchoring.The mammary gland is the lactation organ of mammals,whose main function is the synthesis and secretion of milk components.In the early stage,our research group conducted whole-genome resequencing analysis and verification of serum samples from sheep with four-nipples and sheep with two-nipple,and found that DEPDC7 gene could be used as a candidate gene for four-nipple traits of sheep with four-nipples.However,there are few reports on the regulation of DEPDC7 gene on the mammary gland development of sheep with four-nipples.Therefore,this study analyzed the differential expression m RNA caused by overexpression of DEPDC7 in the breast epithelial cells of sheep with four-nipples,providing new experimental basis for better exploration of the biological function of DEPDC7 gene in the breast epithelial cells of sheep with four-nipples.Sheep with four-nipples in this study in mammary epithelial cells as the research object,by constructing wild type DEPDC7 gene expression vector,sheep with four-nipples transfection in mammary epithelial cells,the absolute quantitative transcriptome sequencing,and clustering analysis,GO and KEGG pathway analysis,in order to clarify DEPDC7 sheep with four-nipples to play in the development of mammary gland epithelial cell biology function.The main results are as follows:1.In this study,the mammary epithelial cells of sheep with four-nipples were successfully purified and cultured.They were identified as positive by immunofluorescence technology and in good condition.The obtained cells can be determined as the mammary epithelial cells of sheep with four-nipples.2.In this study,absolute quantitative transcriptome sequencing technology was used to detect and compare the differentially expressed m RNA caused by overexpression of DEPDC7 in sheep with four-nipples mammary epithelial cells,Screened 116 significantly differentially expressed genes,of which 104 were up-regulated and 12 down-regulated,indicating that the expression levels of these genes are regulated or affected by DEPDC7;3.Analysis of GO and KEGG signal pathways on differentially expressed genes revealed that multiple genes are involved in the growth and development of sheep with four-nipples mammary epithelial cells and differentially expressed,providing a basis for further studying the biological function of DEPDC7 gene.To sum up,this study found that the 116 gene caused by overexpression of DEPDC7 in the breast epithelial cells of sheep with four-nipples was differenciously expressed at the m RNA level,which confirmed that DEPDC7 gene was involved in the growth and development of the breast epithelial cells of sheep with four-nipples and may be involved in the development of the accessory papilla.