DDR1基因过表达对水牛乳腺上皮细胞的影响

doi:10.16431/j.cnki.1671-7236.2019.12.021

Abstract

Abstract: In order to reveal the effect of discoidin domain receptor 1 (DDR1) gene on the lactation performance of buffalo,in this study,the eukaryotic expression vector of buffalo DDR1 gene was constructed,and its optimal transfection time was groped.The effects of DDR1 gene expression on buffalo mammary epithelial cells were also analyzed.Results of agarose gel electrophoresis showed that the fragment size of vetor was 8.8 kb,it was the same as the target carrier fragment size,and the sequencing results showed that the match rate with the target fragment sequence was 100%.Results of cell transfection test showed that the optimal transfection time of DDR1 gene over expression was 48 h.Results of cell proliferation test showed that there was no significant difference in the relative fluorescence value between the DDR1 gene over expression group and the control group (P>0.05).Apoptosis test results showed that the late apoptosis rate was not affected (19.87% VS 17.49%) by the DDR1 gene over expression (P>0.05),but the early apoptosis rate of buffalo mammary epithelial cells (6.48% VS 1.35%) was extremely significantly increased (P<0.01).Moreover,DDR1 gene over expression increased the expression of apoptosis-inhibiting genes BCL-2 and XIAP in buffalo mammary epithelial cells,and reduced the expression of apoptosis-promoting gene P53.In addition,over expression of the DDR1 gene significantly improved the migration rate of buffalo mammary epithelial cells (66.26% VS 58.76%,P<0.05).In conclusion,the research successfully constructed the eukaryotic expression vector of buffalo DDR1 gene and proved that over expression of DDR1 gene promoted the early apoptosis and migration of buffalo mammary epithelial cells,and laid a foundation for the future study of the development and lactation performance of buffalo mammary gland.

Key words: DDR1 gene; buffalo; mammary epithelial cells

CLC Number:

S813.3

LU Xingrong, DUAN Anqin, MA Xiaoya, LIANG Shasha, PANG Chunying, LIANG Xianwei, DENG Tingxian. Effect of DDR1 Gene over Expression on Buffalo Mammary Epithelial Cells[J]. China Animal Husbandry and Veterinary Medicine, 2019, 46(12): 3635-3641.

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Effect of DDR1 Gene over Expression on Buffalo Mammary Epithelial Cells

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