Genetic Evidence for the Involvement of the S-Layer Protein Gene sap and the Sporulation Genes spo0A, spo0B, and spo0F in Phage AP50c Infection of Bacillus anthracis

Roger D. Plaut; John W. Beaber; Jason Zemansky; Ajinder P. Kaur; Matroner George; Biswajit Biswas; Matthew Henry; Kimberly A. Bishop-Lilly; Vishwesh Mokashi; Ryan M. Hannah; Robert K. Pope; Timothy D. Read; Scott Stibitz; Richard Calendar; Shanmuga Sozhamannan

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Genetic Evidence for the Involvement of the S-Layer Protein Gene sap and the Sporulation Genes spo0A, spo0B, and spo0F in Phage AP50c Infection of Bacillus anthracis

机译：S层蛋白基因树液和孢子基因spo0A，spo0B和spo0F参与炭疽杆菌噬菌体AP50c感染的遗传证据。

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In order to better characterize the Bacillus anthracis typing phage AP50c, we designed a genetic screen to identify its bacterial receptor. Insertions of the transposon mariner or targeted deletions of the structural gene for the S-layer protein Sap and the sporulation genes spo0A, spo0B, and spo0F in B. anthracis Sterne resulted in phage resistance with concomitant defects in phage adsorption and infectivity. Electron microscopy of bacteria incubated with AP50c revealed phage particles associated with the surface of bacilli of the Sterne strain but not with the surfaces of Δsap, Δspo0A, Δspo0B, or Δspo0F mutants. The amount of Sap in the S layer of each of the spo0 mutant strains was substantially reduced compared to that of the parent strain, and incubation of AP50c with purified recombinant Sap led to a substantial reduction in phage activity. Phylogenetic analysis based on whole-genome sequences of B. cereus sensu lato strains revealed several closely related B. cereus and B. thuringiensis strains that carry sap genes with very high similarities to the sap gene of B. anthracis. Complementation of the Δsap mutant in trans with the wild-type B. anthracis sap or the sap gene from either of two different B. cereus strains that are sensitive to AP50c infection restored phage sensitivity, and electron microscopy confirmed attachment of phage particles to the surface of each of the complemented strains. Based on these data, we postulate that Sap is involved in AP50c infectivity, most likely acting as the phage receptor, and that the spo0 genes may regulate synthesis of Sap and/or formation of the S layer.

机译：为了更好地表征炭疽芽孢杆菌分型噬菌体AP50c，我们设计了基因筛选来鉴定其细菌受体。 S层蛋白Sap的结构基因以及孢子形成基因 spo0A ， spo0B 和 spo0F 的转座子水手的插入或靶向缺失炭疽芽孢杆菌中的斯特恩导致噬菌体抗性以及噬菌体吸附和感染性的缺陷。用AP50c孵育的细菌的电子显微镜观察到，噬菌体颗粒与Sterne菌株的细菌表面有关，但与Δ sap ，Δ spo0A ，Δ的表面无关spo0B 或Δ spo0F 突变体。与亲本菌株相比，每个 spo0 突变菌株S层中Sap的量均显着减少，并且将AP50c与纯化的重组Sap孵育导致噬菌体活性大大降低。基于蜡状芽孢杆菌全基因组序列的系统发育分析，揭示了数个紧密相关的蜡状芽孢杆菌和苏云金芽孢杆菌菌株，它们携带与 sap 非常相似的 sap 基因。 em>炭疽芽孢杆菌的基因。野生型炭疽芽孢杆菌 sap 或 sap 基因中的 trans 中的Δ sap 突变体的互补对AP50c感染敏感的两种不同蜡状芽孢杆菌菌株均恢复了噬菌体敏感性，电子显微镜证实噬菌体颗粒附着在每个互补菌株的表面。根据这些数据，我们推测Sap参与AP50c的感染性，最有可能充当噬菌体受体，而 spo0 基因可能调节Sap的合成和/或S层的形成。 展开▼

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《Journal of bacteriology》 |2014年第6期|共12页

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Roger D. Plaut; John W. Beaber; Jason Zemansky; Ajinder P. Kaur; Matroner George; Biswajit Biswas; Matthew Henry; Kimberly A. Bishop-Lilly; Vishwesh Mokashi; Ryan M. Hannah; Robert K. Pope; Timothy D. Read; Scott Stibitz; Richard Calendar; Shanmuga Sozhamannan;
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专利

1. Characterization of the gene for a protein kinase which phosphorylates the sporulation-regulatory proteins Spo0A and Spo0F of Bacillus subtilis. [J] . M Perego, S P Cole, D Burbulys, Journal of bacteriology . 1989,第11期

机译：磷酸化枯草芽孢杆菌孢子调节蛋白Spo0A和Spo0F的蛋白激酶基因的表征。

2. Bacillus subtilis early sporulation genes kinA, spo0F, and spo0A are transcribed by the RNA polymerase containing sigma H. [J] . M Predich, G Nair, I Smith Journal of bacteriology . 1992,第9期

机译：枯草芽孢杆菌早期孢子形成基因kinA，spo0F和spo0A被包含sigma H的RNA聚合酶转录。

3. Genetic analysis of Bacillus anthracis Sap S-layer protein crystallization domain [J] . Thomas Candela, Tam Mignot, Xavier Hagnerelle, Microbiology . 2005,第5期

机译：杆菌炭疽病S层蛋白结晶结构域的遗传分析

4. (67a) Host Genetic interactions During Viral Infection: A Systems Virology Approach to Lambda Phage Infection of E. Coli [C] . Elsa Birch1, Nathaniel Maynard2 and Markus Covert American Institute of Chemical Engineers annual meeting . 2010

机译：（67A）病毒感染期间宿主遗传相互作用：大肠杆菌λ噬菌体感染的系统病毒学方法

5. Transcriptional regulation of genes encoding spore coat proteins by mother-cell specific sigma(K) RNA polymerase during Bacillus subtilis sporulation. [D] . Ichikawa, Hiroshi Travis. 2000

机译：枯草芽孢杆菌形成过程中母细胞特异性sigma（K）RNA聚合酶对编码孢子外壳蛋白的基因的转录调控。

6. Genetic Evidence for the Involvement of the S-Layer Protein Gene sap and the Sporulation Genes spo0A spo0B and spo0F in Phage AP50c Infection of Bacillus anthracis [O] . Roger D. Plaut, John W. Beaber, Jason Zemansky, 2014

机译：S层蛋白基因树液和spor0Aspo0B和spo0F基因参与炭疽杆菌噬菌体AP50c感染的遗传证据。

7. Bacillus subtilis early sporulation genes kinA, spo0F, and spo0A are transcribed by the RNA polymerase containing sigma H. [O] . Predich, M, Nair, G, Smith, I 1992

机译：枯草芽孢杆菌早期孢子形成基因kinA，spo0F和spo0A被包含sigma H的RNA聚合酶转录。

8. Genes for Bacillus Megaterium Small, Acid-Soluble Spore Proteins: Nucleotide Sequence of Two Genes and Their Expression during Sporulation [R] . Fliss, E. R., Setlow, P. 1985

机译：巨大芽孢杆菌小，酸可溶性孢子蛋白的基因：两个基因的核苷酸序列及其在孢子形成过程中的表达

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3. 狐源兔脑炎微孢子虫分离鉴定及极管蛋白基因遗传变异分析 [A] . 李志 . 2019

1. ANALYSIS METHOD OF MACROPHAGE REACTION GENE BY BACILLUS ANTHRACIS LETHAL TOXIN TO DETECT INFECTION OF BACILLUS ANTHRACIS [P] . 外国专利： KR20050003815A . 2005-01-12

机译：炭疽芽孢杆菌甲状旁腺毒素检测巨噬细胞反应基因的方法

2. Plasmid containing DNA from a PLASMIDFRAKTION OF B.THURINGIENSIS, E.COLI bacteria strain containing the plasmid, E.COLI genetically engineered bacterial strain, method for preparing the plasmid method of creating a genetically engineered bacterial strain E.COLI, process for producing Bacillus thuringiensis CRYSTAL PROTEINS AND PROCEDURE TO OBTAIN insecticidal activity IN AN ENVIRONMENT [P] . 外国专利： DK160998B . 1991-05-13

机译：包含来自苏云金芽孢杆菌的质粒的DNA的质粒，包含该质粒的大肠杆菌大肠杆菌菌株，大肠杆菌基因工程菌菌株，制备该质粒的方法，用于制备遗传细菌菌株大肠杆菌的方法，生产苏云金芽孢杆菌的方法在环境中的晶体蛋白和获得杀虫活性的方法

3. Genetically engineered hepatitis c virus (hcv) nucleic acid clone, plasmid clone that houses a life-size hcv cdna, dna or rna transcribed from the full-length hcv cdna, transfected and in vitro infected host cell line with hcv, non-human animal infected with hcv, hcv viral particle, in vitro test system, hcv cell free, hcv vaccine and hcv test set and processes for identifying a cell line that is permissive and an animal that is permissive for hcv infection, to select hcv with adaptive mutations that allow higher levels of hcv replication in a permissive cell line, to infect an animal with hcv, to spread hcv in vitro, to transduce an animal susceptible to hcv infection with a heterologous gene, to produce hcv viral particles, hcv particle proteins and hcv antibodies, to separate agents capable of to modulate hcv replication, to prepare a hcv nucleic acid and to detect hcv antibodies in a biological sample from a patient. [P] . 外国专利： BR9808147A . 2000-03-28

机译：基因工程丙型肝炎病毒（hcv）核酸克隆，质粒克隆，其中包含从全长hcv cdna转录而来的真人大小的hcv cdna，dna或rna，并通过非人类动物的hcv转染和体外感染宿主细胞系被hcv感染，hcv病毒颗粒，体外测试系统，无hcv细胞，hcv疫苗和hcv测试仪的感染以及鉴定允许的细胞系和允许感染hvv的动物的过程，以选择具有以下适应性突变的hcv：允许在允许的细胞系中进行更高水平的hvv复制，以hvv感染动物，在体外传播hvv，用异源基因转导易感染hvv的动物，产生hvv病毒颗粒，hvv颗粒蛋白和hvv抗体分离能够调节hcv复制的试剂，制备hcv核酸并检测患者生物样品中的hcv抗体。

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Genetic Evidence for the Involvement of the S-Layer Protein Gene sap and the Sporulation Genes spo0A, spo0B, and spo0F in Phage AP50c Infection of Bacillus anthracis

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