组蛋白甲基化抑制剂UNC0638对水牛转基因细胞外源基因表达的影响

doi:10.11843/j.issn.0366-6964.2018.10.012

Abstract

Abstract:

The purpose of this study was to evaluate the effect of different concentrations of the histone methylation inhibitor UNC0638 on the expression level of transgenic buffalo fibroblast exogenous gene to provide a theoretical basis for studying the mechanism of exogenous gene expression and improving the production efficiency of transgenic buffaloes. After the buffalo fetal fibroblasts (BFFs) were treated with different concentrations of UNC0638(0(control group),0.5,1.5,2.5,5.0 mol·L^-1)for 8 d, the cell growth curve was plotted and the cell karyotype was detected after treated with different concentrations of UNC0638 for 48 h. The methylation level of the histone H3K9me2 of BFFs was detected by immunofluorescent technique, and the effect of UNC0638 on the expression efficiency of buffalo transgenic cells exogenous genes was analyzed by quantitative real-time PCR (qRT-PCR). The results showed that:1) The growth curves of BFFs treated with different concentrations of UNC0638 were similar to that of the control group and all showed "S" type,and 0.5, 1.5, 2.5 μmol·L^-1 UNC0638 had no significant effect on the cell proliferation, while 5.0 μmol·L^-1 UNC0638 inhibited the BFFs proliferation. 2)No significant difference was observed in the normal rate of the cell karyotype among the different groups. 3)In addition,the methylation level of histone H3K9me2 in BFFs treated with UNC0638 was significantly decreased compared with the control group(P<0.05). 4)The qRT-PCR results showed that the relative expression of eGFP in the buffalo transgenic fetal fibroblasts treated with 2.5 μmol·L^-1 UNC0638 was significantly higher than that in the control group in 24 h (P<0.05), while the 0.5, 1.5 and 2.5 μmol·L^-1 UNC0638 groups were significantly higher than that in the control group in 48 h (P<0.05). In conclusion,the research showed that UNC0638 could effectively reduce the methylation level of the buffalo fibroblast histone H3K9me2 and increase the efficiency of exogenous gene expression in buffalo transgenic cells.

CLC Number:

S823.8⁺3.2

LIU Xiao-hua, WEN Dong-mei, YU Qing, DENG Kai, LU Feng-hua, SHI De-shun. Effect of the Histone Methylation Inhibitor UNC0638 on Exogenous Genes Expression of Transgenic Buffalo Fibroblast[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(10): 2163-2169.

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Effect of the Histone Methylation Inhibitor UNC0638 on Exogenous Genes Expression of Transgenic Buffalo Fibroblast

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