牛种布鲁氏菌TLR9_CpG_DNA免疫信号通路活性检测

Evaluation of TLR9_Cp G_DNA Signaling Pathway Activity From Brucella Abortus

目的检测牛种布鲁氏菌TLR9_Cp G_DNA免疫信号通路活性。方法 NCBI网站上下载Brucella abortus A13334全部基因组序列,利用生物信息学的方法筛选Cp G基序,合成天然骨架的脱氧寡核苷酸(ODNs),转染小鼠单核巨噬细胞株RAW264.7收集上清,Elisa检测炎性因子干扰素IFN-α的分泌水平。结果共筛选出1 935条牛种布鲁氏菌Cp G,选其中二级茎环结构较好的5条序列合成ODNs,Elisa结果显示,序列M1分泌的IFN-α浓度与对照序列相比表达量增高差异有统计学意义。结论宿主TLR9识别牛种布鲁氏菌Cp G基序,从而活化免疫信号通路。

Objective The aim of this study was to screen natural DNA sequences named CpG with TLR9 agonistic activity from Brucella abortus. Method Complete genome sequence of Brucella abortus A13334 was available in genebank. Bioinformatics technology was used to identify Brucella abortus CpG. Phosphodiester ODNs representing the selected ODNs were then synthesized. Murine macrophage RAW264.7 were cultured and transfected with ODNs. The content of IFN - a in the supernatant was then measured by ELISA. Result Totally screened 1 935 CpG in abortus brucella, 5 CpG sequences was synthesized to ODNs. Elisa results showed compared with the control sequence, the difference of Concentration of IFN - α in RAW264.7 cells originate from sequence M1 was statistically increased. Conclusion This work demonstrated that CpG sequences present in Brucella abortus DNA were able to produce macrophages stimulation through TLR9.