A group B Streptococcus indexed transposon mutant library to accelerate genetic research on an important perinatal pathogen.
A group B Streptococcus indexed transposon mutant library to accelerate genetic research on an important perinatal pathogen.
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DOI：
10.1128/spectrum.02046-23
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发表时间：
2023-12-12
影响因子：
3.7
通讯作者：
Hooven, Thomas A.
中科院分区：
文献类型：
--
作者：
Bhavana, Venkata H.;Hillebrand, Gideon H.;Gopalakrishna, Kathyayini P.;Rapp, Rebekah A.;Ratner, Adam J.;Tettelin, Herve;Hooven, Thomas A.
B群链球菌（GBS）是新生儿败血症、脑膜炎和肺炎的主要致病因素。通过允许快速筛选导致疾病的基因，索引细菌突变文库加速了发病机制的研究。在这项研究中，我们从一个混合转座子插入文库中创建了一个大规模的GBS索引的Himar1迷你转座子突变菌株文库，该文库是我们之前用于转座子-基因组连接测序的。我们使用高通量工作流程来鉴定从单个突变体纯化的染色体DNA中的转座子插入位点。在质量控制步骤和去除等基因重复菌株之后，我们分离出1919个单培养的独特转座子插入突变体，这些突变体在GBS基因组中分布均匀。我们最终的文库，储存在条形码，可追溯的甘油库存中，包含878个基因的中断和253个基因间区域。我们还用验证性PCR验证了选择的文库突变体，并在可能的情况下进行了特异性表型检测。虽然文库只包含稀疏的必要或近必要基因的中断，但在集合中代表的基因跨越了广泛的预测功能类别，包括代谢，结构和毒力中的作用。总之，我们开发并应用了高通量分子分析和生物信息学管道来生成一个规模空前的GBS索引库，我们相信这将成为GBS研究人员的有用遗传工具。B群链球菌（GBS）是全球严重感染的重要原因，其中大多数影响孕妇，新生儿和婴儿。研究GBS基因突变菌株是了解这些感染是如何引起的一个有价值的方法，也是开发更有效的预防和治疗策略的关键一步。在这个资源报告中，我们描述了一个新创建的定义GBS基因突变文库，包含超过1900个遗传变异，每个变异都有一个独特的染色体断裂。如此规模的索引库在GBS领域是前所未有的；它包括数百种基因突变的菌株，这些基因在人类疾病中的潜在功能尚不清楚。我们通过在公共资助的细菌维护和分布库中沉积，使该资源免费提供给更广泛的研究界。
Group B Streptococcus (GBS) is a major contributor to sepsis, meningitis, and pneumonia in newborns. Indexed bacterial mutant libraries accelerate pathogenesis research by allowing rapid screening of genes that contribute to disease. In this study, we created and characterized a large-scale GBS indexed library of Himar1 mini-transposon mutant strains grown as monocultures from a mixed transposon insertion library that we had previously used for transposon-genome junction sequencing. We used a high-throughput workflow to identify transposon insertion sites in chromosomal DNA purified from individual mutants. Following quality control steps and the removal of isogenic duplicate strains, we isolated 1,919 monocultures of unique transposon insertion mutants with even dispersion across the GBS genome. Our final library, stored in barcoded, traceable glycerol stocks, contains interruptions of 878 genes and 253 intergenic regions. We also validated select library mutants with confirmatory PCR and, when possible, specific phenotypic testing. While the library contains only sparse interruptions of essential or near-essential genes, the genes represented in the set span a wide range of predicted functional categories, including roles in metabolism, structure, and virulence. In conclusion, we developed and applied a high-throughput molecular analysis and bioinformatic pipeline to generate a GBS indexed library of unprecedented scale that we believe will be a useful genetic tool for fellow GBS researchers. Group B Streptococcus (GBS) is a significant global cause of serious infections, most of which affect pregnant women, newborns, and infants. Studying GBS genetic mutant strains is a valuable approach for learning more about how these infections are caused and is a key step toward developing more effective preventative and treatment strategies. In this resource report, we describe a newly created library of defined GBS genetic mutants, containing over 1,900 genetic variants, each with a unique disruption to its chromosome. An indexed library of this scale is unprecedented in the GBS field; it includes strains with mutations in hundreds of genes whose potential functions in human disease remain unknown. We have made this resource freely available to the broader research community through deposition in a publicly funded bacterial maintenance and distribution repository.
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影响因子：
3.7
作者：
通讯作者：
--
影响因子：
6.4
作者：
Dale JL;Beckman KB;Willett JLE;Nilson JL;Palani NP;Baller JA;Hauge A;Gohl DM;Erickson R;Manias DA;Sadowsky MJ;Dunny GM
通讯作者：
Dunny GM
影响因子：
9.9
作者：
de Berardinis, Veronique;Vallenet, David;Castelli, Vanina;Besnard, Marielle;Pinet, Agnes;Cruaud, Corinne;Samair, Sumitta;Lechaplais, Christophe;Gyapay, Gabor;Richez, Celine;Durot, Maxime;Kreimeyer, Annett;Le Fevre, Francois;Schaechter, Vincent;Pezo, Valerie;Doering, Volker;Scarpelli, Claude;Medigue, Claudine;Cohen, Georges N.;Marliere, Philippe;Salanoubat, Marcel;Weissenbach, Jean
通讯作者：
Weissenbach, Jean
影响因子：
14.9
作者：
Chen, I-Min A.;Chu, Ken;Kyrpides, Nikos C.
通讯作者：
Kyrpides, Nikos C.
影响因子：
3.1
作者：
BROWN, J;FARNSWORTH, R;JOHNSON, DW
通讯作者：
JOHNSON, DW