| 摘要: |
| 为研究羊种布鲁氏菌转录调控因子MucR蛋白的毒力相关机制,对染色质免疫共沉淀技术进行了优化。构建了表达MucRFlag融合蛋白的羊种布鲁氏菌菌株,并用商品化抗Flag标签抗体对MucRFlag蛋白进行富集,从而替代MucR蛋白抗体的制备过程。通过该染色质免疫共沉淀方法对羊种布鲁氏菌MucR蛋白的结合靶点基因进行了鉴定,结果显示:通过使用该方法鉴定出羊种布鲁氏菌MucR蛋白可以结合到BMEI0430和BMEI1364(mucR基因)的启动子序列上。结果表明优化的染色质免疫共沉淀技术适用布鲁氏菌转录调控因子结合靶点的研究。 |
| 关键词: 布鲁氏菌 染色质免疫共沉淀 转录调控因子 靶点基因 |
| DOI：10.11841/j.issn.1007-4333.2016.04.13 |
| 投稿时间：2015-06-11 |
| 基金项目:国家973项目(2010CB530202);国家自然科学基金资助项目(30871882) |
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| Identification of the targets of MucR by chromatin immunoprecipitation in Brucella melitensis |
|
DONG Hao1,2, PENG Xiao-wei1, WANG Xiao-ying2, SUN Yu2, SONG Xiao-hui2, WU Qing-min1
|
| (1.College of Veterinary Medicine, China Agriculture University, Beijing 100193, China;2.China Animal Disease Control Centre, Beijing 102600, China) |
| Abstract: |
| To investigate virulence-related mechanism of MucR in Brucella melitensis, the chromatin immunoprecipitation technology was optimized.A MucRFlag protein expression strain was constructed, and its fusion proteins were enriched by commercial anti-Flag tag antibody with less timeconsuming and expense compared to preparing and using MucR antibody.In this study, we used optimized chromatin immunoprecipitation method to identify targets of MucR protein.Promoter sequences of two genes (BMEI0430 and BMEI1364), which could bind to MucR protein in B.melitensis, were identified.These results demonstrated that the method cannot only be applied in identification the targets of transcription regulators in Brucella spp., but also provide potential strategy for the study targets genes in other bacterial transcriptional regulatory networks. |
| Key words: Brucella melitensis chromatin immunoprecipitation transcriptional regulators target genes |
