Shuttle vectors for Chlamydia trachomatis

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1 Supplemental Material: Annu. Rev. Microbiol : doi: /annurev-micro Molecular Genetic Analysis of Chlamydia Species Sixt and Valdivia Supplemental Table 1: Shuttle vectors for Chlamydia Vector name ppbw100 pbr325::l2 pgfp::sw2 pgfp-cat::sw2 IncDProm-RSGFP- IncDTerm IncDPrommCherry-IncDTerm IncDProm-CFP- IncDTerm pcte1 (C. trachomatis E/UW-5/CX) CDS1 and CDS3 L2/434/Bu) CDS1 Shuttle vectors for Chlamydia trachomatis (Kanamycin) none none First successful but transient transformation of Chlamydia First stable transformation of Chlamydia First stable transformation of Chlamydia with a vector conferring expression of a fluorescent protein Derivative of pgfp::sw2 lacking ß-lactamase gene; first description of chloramphenicol for selection in Chlamydia NA b (12) (16) (16) (19) none 9184 (1) rsgfp (C. trachomatis incd promoter and mcherry (C. trachomatis incd promoter and CFP (C. trachomatis incd promoter and Derivative of expressing rsgfp Derivative of expressing mcherry Derivative of expressing CFP (1) (1) (1)

2 pbrct pbra pgfpbsd/z::sw2 pask-gfp-l2 pask- GFP/mKate2-L2 ptl2-tap puc-pcl2 pdnak-rogfp2- SW2 pbomb4 pbomb4-mci L2/434/Bu) all ORFs pcta (C. trachomatis A2497) all ORFs E/SW2) CDS1 E/SW2) CDS1 none (10) none Derivative of pbrct containing from C. trachomatis A Blasticidin (Zeocin) GFP-BSD GFP (teta promoter, inducible) GFP (teta promoter, inducible); mkate2 (weakened C. trachomatis dnak First description of blasticidin S for selection in Chlamydia First description of inducible gene expression in Chlamydia Derivative of pask-gfp-l2 with constitutive expression of mkate2 none Derivative of pask-gfp-l2 for inducible expression of TAPtagged proteins none First description of dendrimerenabled transformation of C. trachomatis rogfp2 (C. trachomatis dnak rsgpp mcherry Designed to monitor the redox state in the cytosol of Chlamydia MCS containing BamHI, SacII, NotI, NheI, PstI, AgeI, KpnI, and SalI Derivative of pbomb4 expressing mcherry (11) NA b (5) (18) (18) NA b (2) NA b (7) NA b (14) (3) NA b (3)

3 pbomb4r rsgfp Derivative of pbomb4 with C. trachomatis rpob promoter upstream of MCS (3) pbomb4r-mci pbomb4-tetmcherry pbomb3 pct308-gfp pl2dest mcherry rsgfp ; mcherry (tetracycline inducible (tetracycline inducible mcherry Derivative of pbomb4-mcl with C. trachomatis rpob promoter upstream of MCS Derivative of pbomb4 with tetracycline inducible promoter in MCS NA b (3) (3) Derivative of pbomb (17) (13) (9) Vector name pbrcm pcm (C. muridarum) all ORFs Shuttle vectors for Chlamydia muridarum none Derivative of pbrct containing from C. muridarum (11)

4 pgfp::cm pcm (C. muridarum Nigg) all ORFs Derivative of pgfp::sw2 containing from C. muridarum NA b (8) pgfp-luc-cm psw2niggcds2 pcm (C. muridarum Nigg) all ORFs ; CDS2 replaced by CDS2 from C. muridarum pnigg ; Luciferase (C. muridarum pgp4 promoter, C. trachomatis D CT579 Derivative of pgfp::cm expressing luciferase Originates from recombination between pgfp::sw2 and the endogenous from C. muridarum Nigg NA b (4) (15) Vector name pcl2-eg(kg) Shuttle vectors for Chlamydia pneumoniae (URA3, LEU2-d) none Chlamydia-yeast shuttle vector; no antibiotic selection marker for Chlamydia, first description of dendrimer-enabled transformation of Chlamydia with a shuttle vector NA b (6) a Selection markers in parentheses were used for selection in E.coli or S. cerevisiae, but not Chlamydia b NA refers to not available (information not given in the reference)

5 References 1. Agaisse H, Derre I A C. trachomatis cloning vector and the generation of C. trachomatis strains expressing fluorescent proteins under the control of a C. trachomatis promoter. PLoS One 8:e Barta ML, Battaile KP, Lovell S, Hefty PS Hypothetical protein CT398 (CdsZ) interacts with sigma (RpoN)-holoenzyme and the type III secretion export apparatus in Chlamydia trachomatis. Protein Sci 24: Bauler LD, Hackstadt T Expression and targeting of secreted proteins from Chlamydia trachomatis. J Bacteriol 196: Campbell J, Huang Y, Liu Y, Schenken R, Arulanandam B, Zhong G Bioluminescence imaging of Chlamydia muridarum ascending infection in mice. PLoS One 9:e Ding H, Gong S, Tian Y, Yang Z, Brunham R, Zhong G Transformation of sexually transmitted infection-causing serovars of Chlamydia trachomatis using blasticidin for selection. PLoS One 8:e Gérard HC, Mishra MK, Mao G, Wang S, Hali M, et al Dendrimer-enabled DNA delivery and transformation of Chlamydia pneumoniae. Nanomedicine 9: Kannan RM, Gerard HC, Mishra MK, Mao G, Wang S, et al Dendrimer-enabled transformation of Chlamydia trachomatis. Microb Pathog 65: Liu Y, Chen C, Gong S, Hou S, Qi M, et al Transformation of Chlamydia muridarum reveals a role for Pgp5 in suppression of -dependent gene expression. J Bacteriol 196: Mueller KE, Fields KA Application of beta-lactamase reporter fusions as an indicator of effector protein secretion during infections with the obligate intracellular pathogen Chlamydia trachomatis. PLoS One 10:e Song L, Carlson JH, Whitmire WM, Kari L, Virtaneva K, et al Chlamydia trachomatis -encoded Pgp4 is a transcriptional regulator of virulence-associated genes. Infect Immun 81: Song L, Carlson JH, Zhou B, Virtaneva K, Whitmire WM, et al Plasmid-mediated transformation tropism of chlamydial biovars. Pathog Dis 70: Tam JE, Davis CH, Wyrick PB Expression of recombinant DNA introduced into Chlamydia trachomatis by electroporation. Can J Microbiol 40: Thompson CC, Griffiths C, Nicod SS, Lowden NM, Wigneshweraraj S, et al The Rsb phosphoregulatory network controls availability of the primary sigma factor in Chlamydia trachomatis and influences the kinetics of growth and development. PLoS Pathog 11:e Wang X, Schwarzer C, Hybiske K, Machen TE, Stephens RS Developmental stage oxidoreductive states of Chlamydia and infected host cells. MBio 5:e Wang Y, Cutcliffe LT, Skilton RJ, Ramsey KH, Thomson NR, Clarke IN The genetic basis of tropism between Chlamydia trachomatis and Chlamydia muridarum. Pathog Dis 72: Wang Y, Kahane S, Cutcliffe LT, Skilton RJ, Lambden PR, Clarke IN Development of a transformation system for Chlamydia trachomatis: restoration of glycogen biosynthesis by acquisition of a shuttle vector. PLoS Pathog 7:e Weber MM, Noriea NF, Bauler LD, Lam JL, Sager J, et al A functional core of IncA is required for Chlamydia trachomatis inclusion fusion. J Bacteriol 198: Wickstrum J, Sammons LR, Restivo KN, Hefty PS Conditional gene expression in Chlamydia trachomatis using the tet system. PLoS One 8:e Xu S, Battaglia L, Bao X, Fan H acetyltransferase as a selection marker for chlamydial transformation. BMC Res Notes 6:377

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