Chloramphenicol acetyltransferase as a selection marker for chlamydial transformation

BMC Research Notes

Huizhou Fan²  Xiaofeng Bao¹  Lauren Battaglia³  Shuang Xu² 

[1] Department of Pharmacology, Nantong University School of Medicine, Nantong, China;Department of Pharmacology, Rutgers University Robert Wood Johnson Medical School, Piscataway, NJ, USA;Rutgers University Graduate Program in Cell and Developmental Biology, Piscataway, NJ, USA

关键词: Transformation selection marker;    Transformation;    Sexually transmitted infections;    Chloramphenicol acetyltransferase;    Chloramphenicol;    Chlamydia trachomatis;    Chlamydia;   

Others  :  1141602
DOI  :  10.1186/1756-0500-6-377

 received in 2013-07-08, accepted in 2013-09-11,  发布年份 2013

【 摘 要 】

Background

Chlamydia is a common bacterial pathogen responsible for many diseases. Methods for transforming this important organism using a β-lactamase as a selection marker have been developed very recently. However, the National Institutes of Health Guidelines for Research Involving Recombinant DNA Molecules do not permit transformation experiments with β-lactamase gene-containing vectors for certain human chlamydial pathogens. Therefore, a different selection marker is urgently needed for transformation of those chlamydiae.

Results

After transformation of plasmid-free Chlamydia trachomatis with pGFP:SW2, which carries a β-lactamase and a chloramphenicol acetyltransferase gene fused to a green fluorescence protein gene, transformants were obtained by selection with either ampicillin or chloramphenicol. Stable chloramphenicol-resistant, but ampicillin-sensitive, transformants were obtained using a pGFP:SW2 derivative without the β-lactamase. All transformants expressed green fluorescence protein and had glycogen synthesis activity restored.

Conclusions

Chloramphenicol resistance may be used as a selection marker for genetic experiments in Chlamydia. This eliminates the requirement for the use of β-lactamase, of which dissemination to some C. trachomatis serovars may jeopardize clinical treatment of chlamydial infections in pregnant women. Chloramphenicol acetyltransferase may also serve as a useful secondary selection marker for genetic analyses in β-lactamase-transformed chlamydial strains.

【 授权许可】

   
2013 Xu et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

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