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Proteus
Published in Dongyou Liu, Laboratory Models for Foodborne Infections, 2017
Paola Scavone, Victoria Iribarnegaray, Pablo Zunino
Signature-tagged mutagenesis (STM), described by Hensel et al. in 1995, was designed to detect new virulence genes of the target organism Salmonella typhimurium in a murine model of typhoid fever. This technique is a negative selection method in which unique identification tags allow analysis of pools of mutants in mixed populations. In STM, each mutant is tagged with a unique DNA sequence, these allow for coamplification of the DNA tags of a mixed population of mutants by a single polymerase chain reaction (PCR) [56,57]. The tag consists of a short DNA sequence of 40 bp, inserted in a transposon system. DNA tags can be included during allelic replacement (signature-tagged allele replacement) on a systematic genome-wide scale [58]. Detection of the signature tags could be done by dot blot, PCR, or polymorphic tag-length transposon mutagenesis.
Transposon mutagenesis in oral streptococcus
Published in Journal of Oral Microbiology, 2022
Yixin Zhang, Zhengyi Li, Xin Xu, Xian Peng
Transposon mutagenesis is an effective forward genetic strategy for studying gene function by observing the phenotypic changes in mutated genes. Random mutants in a variety of prokaryotes have been created by using different transposon genes such as Tn3 derivatives, IS (insertion sequence) elements, Tn7, Tn5, and mariner. Since the advent of genome sequencing, techniques such as genetic footprinting, signature-tagged Mutagenesis (STM), transposon site hybridization (TraSH), and scanning Linker mutagenesis (SLM) have been developed [17]. And with the advent of next-generation sequencing (NGS), transposon insertion sequencing (TIS) combines it with large-scale transposon insertion mutations to evaluate the essentiality of genetic features and fitness contribution in the bacterial genome in the saturated random mutant libraries. The four TIS techniques published in 2009 include insertion sequencing (INSeq) in Bacteroides thetaiotaomicron [18], high-throughput insertion tracking by deep sequencing (HITS) in Haemophilus influenzae [19], transposon sequencing (Tn-Seq) in S. pneumoniae [20], and transposon-directed insertion site sequencing (TraDIS) in S. Typhi [21]. Those techniques have been widely used in various bacteria to study fitness and virulence, including Enterococcus faecalis [22], Vibrio parahaemolyticus [23], Salmonella enteritidis [24], Edwardsiella piscicida [25], Ralstonia solanacearum [26] and Pantoea [27]. Ultimately, TIS is a key tool for interpreting the rapidly increasing amount of genome sequencing data and is expected to shed light on the function of individual genome features. With the development of transposon technology, TIS has been reviewed from the perspectives of design and analysis [28,29]. Cain et al. discussed recent applications of TIS in answering general biological questions [30]. The present review focuses on oral microorganisms and highlights the application of transposon mutagenesis, including TIS, to oral streptococci, as well as research progress, aiming to better understand the relationship between oral streptococcal phenotype and genotype, which can help clarify the processes of colonization, virulence, and persistence and provides a more reliable basis for investigating relationships with host ecology and disease status. Table 1 and Figure 1 show some articles and conclusions regarding transposon mutagenesis applied to oral streptococci.