鼠伤寒沙门菌小RNA GcvB靶基因的预测与验证

doi:10.11843/j.issn.0366-6964.2020.04.027

畜牧兽医学报 ›› 2020, Vol. 51 ›› Issue (4): 894-898.doi: 10.11843/j.issn.0366-6964.2020.04.027

• 研究简报 • 上一篇

鼠伤寒沙门菌小RNA GcvB靶基因的预测与验证

潘永^1,2, 刘丽娟⁵, 杨阳^1,2, 李晨⁴, 马光强⁶, 杨琦^1,2,3*

1. 贵州大学动物科学学院, 贵阳 550025;
2. 贵州大学动物疫病研究所, 贵阳 550025;
3. 贵州省动物疫病与兽医公共卫生重点实验室, 贵阳 550025;
4. 贵州省畜禽资源遗传管理站, 贵阳 550025;
5. 都匀市农业农村局, 都匀 558000;
6. 黔南民族职业技术学院, 都匀 558000

收稿日期:2019-11-01 出版日期:2020-04-25 发布日期:2020-04-21
通讯作者: 杨琦,主要从事病原微生物学研究,E-mail:yangqinmg@163.com
作者简介:潘永(1997-),男,贵州水城人,硕士生,主要从事病原微生物学研究
基金资助:
国家自然科学基金（31760740；31602065）；贵州省科技厅联合资金项目贵州大学2017年度学术新苗培养及创新探索专项（黔科合LH字［2017］7263；黔科合平台人才［2017］5788）；贵州省科技厅基金（黔科合基础［2016］1047）；贵州省研究生教育创新计划项目（GZZ2017002）

Prediction and Validation of Small RNA GcvB Target Gene of Salmonella Typhimurium

PAN Yong^1,2, LIU Lijuan⁵, YANG Yang^1,2, LI Chen⁴, MA Guangqiang⁶, YANG Qi^1,2,3*

1. College of Animal Science, Guizhou University, Guiyang 550025, China;
2. Institute of Animal Diseases, Guizhou University, Guiyang 550025, China;
3. Guizhou Key Laboratory of Animal Diseases and Veterinary Public Health, Guiyang 550025, China;
4. Guizhou Province Livestock and Poultry Resource Genetic Management Station, Guiyang 550025, China;
5. Agriculture and Rural Affairs Bureau of Duyun City, Duyun 558000, China;
6. Qiannan National Vocational and Technical College, Duyun 558000, China

Received:2019-11-01 Online:2020-04-25 Published:2020-04-21

摘要/Abstract

摘要： gcvB基因敲除株的转录组测序结果注释到KEGG数据库以及GO富集分析，并利用荧光定量PCR对预测的靶基因进行验证。TargetRNA2的预测结果显示，杂交能量最高的narY、yeaK、trpB和STM2732基因均能够和小RNA GcvB产生至少7个连续的碱基互补，narY、yeaK和trpB基因分别与鼠伤寒沙门菌无氧呼吸、脯氨酸与tRNA的识别以及色氨酸的合成相关。荧光定量PCR检测结果显示，narY、yeaK、trpB和STM2732基因在gcvB基因敲除条件下转录水平分别上调3.4、9.8、12.1和18.37倍。以上结果表明，narY、yeaK、trpB和STM2732基因受小RNA GcvB的负调控且可能为直接负调控。本研究为进一步探明小RNA GcvB与靶基因相互作用、小RNA的调控机制以及沙门菌致病机制奠定了基础。

关键词: 沙门菌, 小RNA GcvB, 转录组测序, 靶基因

Abstract: In order to screen the target genes regulated by Salmonella typhimurium small RNA GcvB, this study predicted genes that can interact with GcvB R1, R2 and R3 regions by TargetRNA2 software based on the transcriptome sequencing results of Salmonella typhimurium gcvB knockout strain, annotated the KEGG database and GO enrichment analysis and validated the predicted genes using real-time PCR. The predicted results of Target RNA2 showed that the narY, yeaK, trpB and STM2732 genes with the highest hybridization energy were able to produce at least 7 consecutive base complementary to the small RNA GcvB. NarY, YeaK and TrpB were respectively related to anaerobic respiration, tRNA that recognizes proline, and synthesis of tryptophan. The results of real-time PCR showed that the transcript levels of narY, yeaK, trpB and STM2732 were up-regulated by 3.4, 9.8, 12.1 and 18.37 times, respectively, under gcvB gene knockout conditions. These results indicated that the narY, yeaK, trpB and STM2732 genes were negatively regulated by small RNA GcvB and may be directly negatively regulated. This study laid the foundation for further understanding of the interaction between small RNA GcvB and target genes, the regulation mechanism of small RNA, and the pathogenic mechanism of Salmonella.

Key words: Salmonella, small RNA GcvB, transcriptome sequencing, target genes

中图分类号:

潘永, 刘丽娟, 杨阳, 李晨, 马光强, 杨琦. 鼠伤寒沙门菌小RNA GcvB靶基因的预测与验证[J]. 畜牧兽医学报, 2020, 51(4): 894-898.

PAN Yong, LIU Lijuan, YANG Yang, LI Chen, MA Guangqiang, YANG Qi. Prediction and Validation of Small RNA GcvB Target Gene of Salmonella Typhimurium[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(4): 894-898.

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鼠伤寒沙门菌小RNA GcvB靶基因的预测与验证

Prediction and Validation of Small RNA GcvB Target Gene of Salmonella Typhimurium

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