摘要
目的:构建类鼻疽伯克霍尔德菌(burkholderia pseudomallei,B. pseudomallei)sRNA基因敲除菌株,并对其生物学功能进行初步评价。方法:设计合成引物9sF/9sR、9xF/9xR、R1/F1,扩增sRNA基因上下游同源臂片段,通过酶切、连接和转化,将目的片段克隆至质粒TPR-pK18mobSacB上,运用同源重组的方法获得类鼻疽伯克霍尔德菌sRNA敲除菌株。结果:类鼻疽伯克霍尔德菌敲除株△sRNA构建成功。与野生株HNBP001比较,△sRNA生长速度、泳动能力、生物被膜形成能力均下降,药敏结果无差异。结论:成功构建类鼻疽伯克霍尔德菌sRNA基因敲除株,为进一步研究sRNA在类鼻疽伯克霍尔德菌中的调控机制提供了基础。
Objective:Construction of Burkholderia pseudomallei(B.pseudomallei)sRNA knockout strains and observation of their biological function.Methods:Design 9sF/9sR,9xF/9xR and R1/F1 primers,which were used to amplify the homologous arm fragment upstream and downstream of the sRNA gene,through enzyme cutting,ligation,and transformation,the sRNA gene was knocked out from the B.pseudomallei by homologous recombination method.Results:The sRNA mutant was successfully constructed.In comparison with wild strain HNBP001,the growth rate,motility and biofilm formation ofΔsRNA decreased,but the antibiotic sensitivity has no differences.Conclusion:The sRNA knockout strain of B.pseudomallei was successfully constructed,laying a foundation for further research on its mechanism of regulating B.pseudomallei.
作者
宋心怡
厉安洋
李艳梅
徐淑慧
夏乾峰
SONG Xin-yi;LI An-yang;LI Yan-mei;XU Shu-hui;XIA Qian-feng(Hainan Medical University International School of Public Health and One Health,Haikou 571199,China;Laboratory of Tropical Biomedicine and Biotechnology,School of Tropical Medicine and Laboratory Medicine,Hainan Medical University,Haikou 571199,China)
出处
《海南医学院学报》
CAS
2023年第9期641-646,共6页
Journal of Hainan Medical University
基金
国家自然科学基金(81960002)。
