Abstract：
:Lack of a system for genetic manipulation of Chlamydia trachomatis has been a key challenge to advancing understanding the molecular genetic basis of virulence for this bacterial pathogen. We developed a non-viral, dendrimer-enabled system for transformation of this organism and used it to characterize the effects of inserting the common 7.5 kbp chlamydial plasmid into strain L2(25667R), a C. trachomatis isolate lacking it. The plasmid was cloned in pUC19 and the clone complexed to polyamidoamine dendrimers, producing ∼83 nm spherical particles. Nearly confluent McCoy cell cultures were infected with L2(25667R) and reference strain L2(434). At 16 h post-infection, medium was replaced with dendrimer-plasmid complexes in medium lacking additives (L2(25667R)) or with additive-free medium alone (L2(434)). Three h later complexes/buffer were removed, and medium was replaced; cultures were harvested at various times post-transformation for analyses. Real time PCR and RT-PCR of nucleic acids from transformed cultures demonstrated plasmid replication and gene expression. A previous report indicated that one or more plasmid-encoded product govern(s) transcription of the glycogen synthase gene (glgA) in standard strains. In L2(25667R) the gene is not expressed, but transformants of that strain given the cloned chlamydial plasmid increase glgA expression, as does L2(434). The cloned plasmid is retained, replicated, and expressed in transformants over at least 5 passages, and GFP is expressed when transformed into growing L2(25667R). This transformation system will allow study of chlamydial gene function in pathogenesis.
journal_name
Microb Pathogjournal_title
Microbial pathogenesisauthors
Kannan RM,Gérard HC,Mishra MK,Mao G,Wang S,Hali M,Whittum-Hudson JA,Hudson APdoi
10.1016/j.micpath.2013.08.003subject
Has Abstractpub_date
2013-12-01 00:00:00pages
29-35eissn
0882-4010issn
1096-1208pii
S0882-4010(13)00116-2journal_volume
65pub_type
杂志文章abstract：:In the present study, we isolated Lactobacillus sakei strain DGH5 from raw beef meat. This bacterium plays an inhibitory effect against food-spoiling bacteria and food-borne pathogens, including Listeria monocytogenes, a gram-positive and pathogenic bacterium. Lactobacillus sakei strain DGH5 was identified through bot...
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journal_title：Microbial pathogenesis
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journal_title：Microbial pathogenesis
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journal_title：Microbial pathogenesis
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journal_title：Microbial pathogenesis
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journal_title：Microbial pathogenesis
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journal_title：Microbial pathogenesis
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doi：10.1016/j.micpath.2017.12.040
更新日期：2018-02-01 00:00:00
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journal_title：Microbial pathogenesis
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doi：10.1006/mpat.1997.0147
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更新日期：2020-01-01 00:00:00
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journal_title：Microbial pathogenesis
pub_type： 杂志文章
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更新日期：2020-12-01 00:00:00
