| 贾仲昕，赵佳男，季 芳，王 雪，李 刚，王承民，秦建华.芽孢杆菌蛋白酶基因的比较基因组学分析[J].广东农业科学,2022,49(5):110-117 |
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芽孢杆菌蛋白酶基因的比较基因组学分析 |
| Comparative Genomics Analysis ofProtease Genes in Bacillus sp. |
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| DOI：10.16768/j.issn.1004-874X.2022.05.013 |
| 中文关键词: 蛋白酶 芽孢杆菌 比较基因组学 荧光定量 PCR aprX 基因 |
| 英文关键词: protease Bacillus comparative genomics fluorescence quantitative PCR aprX gene |
| 基金项目:河北省现代农业产业技术体系奶牛创新团队建设项目（HBCT2018120205）；河北省重点研发项目（20326603D）；广东省科学院人才专项（2016GDASRC-0205）；北京动物园圈养野生动物技术北京市重点实验室开放课题（ZDK202105） |
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| 中文摘要: |
| 【目的】研究芽孢杆菌产蛋白酶能力与携带的蛋白酶基因的关系，探寻影响芽孢杆菌产蛋白酶能
力的重要基因。【方法】通过 筛选培养基筛选能够产生蛋白酶的菌株，采用福林酚法 检测菌株产生的蛋白酶的活
性，对 15 株芽孢杆菌分离株进行全基因组测序，获得菌株染色体上携带的蛋白酶基因情况并将其分类；采用实
时荧光定量 PCR 方法检测蛋白酶基因 mRNA 的转录水平；通过比较基因组学和细菌全基因组关联分析（BGWAS），
探讨影响细菌产蛋白酶能力高低的原因。【结果】根据菌株染色体上携带的蛋白酶基因情况，可分为 7 种基因
型（G1~G7）。比较基因组学和 BGWAS 分析发现，细菌染色体上携带的蛋白酶基因种类的多寡与其产蛋白酶能
力的高低有直接关系，且缺少 aprX 和 aprE 基因菌株的产蛋白酶能力明显下降。进一步研究基因 mRNA 水平发现，
芽孢杆菌产蛋白酶能力的差异与 aprX 的表达水平有关。【结论】 菌株染色体上携带的蛋白酶基因情况与菌株产
蛋白酶能力之间存在关联。aprX 是影响芽孢杆菌属细菌产胞外蛋白酶能力的重要基因。 |
| 英文摘要: |
| 【Objective】The study aims to explore the relationship between the protease-producing capacity of
Bacillus and the protease genes carried by the bacteria and analyze the important enzyme genes that affect the proteaseproducing capacity of Bacillus.【Method】The protease-producing strain was screened by using the selective medium.
The protease activity of the strains was determined by the Folin method. The protease genes carried on the chromosomes of
the strains were obtained and classified by sequencing the whole genome of 15 strains of Bacillus. The transcription level of
protease gene mRNA was measured by the real-time fluorescent quantitative PCR method. Through comparative genomics and Bacterial Genome-Wide Association Analysis (BGWAS), the reasons that affect the ability of bacteria to produce protease
were explored. 【Result】According to the protease genes carried on the chromosomes of the strains, they could be divided
into 7 groups of genotypes (G1-G7). Comparative genomics and BGWAS found that the number of protease genes carried on
bacterial chromosomes was directly related to the protease production capacity, and the protease production capacity of strains
lacking aprX and aprE was significantly reduced. The difference in the protease production capacity of Bacillus was found to
be related to the expression level of aprX by further studying the mRNA level of the gene.【Conclusion】There is a correlation
between the protease gene carried on the chromosome of the strain and the protease production capacity of the strain. The
important enzyme gene that affects the protease production capacity of Bacillus is aprX. |
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