Characterization of the Basic Replicon of Rhodococcus Plasmid PSOX and Development of a Rhodococcus-Escherichia Coli Shuttle Vector

Overview

Journal Appl Environ Microbiol

Specialties Environmental Health
Microbiology

Date 1998 Oct 31

PMID 9797291

Citations 3

Authors

H Bergeron

D Labbe

C W Greer

J Hawari

M J Grossman

B M Sankey

P C Lau

Affiliations

Soon will be listed here.

Abstract

The replication region of a 100-kb desulfurization plasmid (pSOX) from Rhodococcus sp. strain X309 was localized to a 4-kb KpnI fragment, and its sequence was determined. The amino acid sequence of one of the predicted open reading frames (ORFs) was related to the putative replication (Rep) protein sequences of the mycobacterial pLR7 family of plasmids. Three of the five predicted ORF products were identified by radiolabelling with the Escherichia coli T7 polymerase/promoter system. In E. coli, the Rep protein of pSOX was apparently synthesized in a shortened form, 21.3 kDa instead of the predicted 41.3 kDa, as a result of an internal initiation. This situation is reminescent of that for some bacterial Rep proteins. A shuttle plasmid was constructed with the pSOX origin, pBluescript II KS-, and the chloramphenicol resistance (Cmr) gene from pRF29. This new shuttle plasmid was used to demonstrate expression of the Bacillus subtilis sacB gene in a strain of Rhodococcus, rendering it sensitive to the presence of sucrose.

Citing Articles

Functional characterization of a catabolic plasmid from polychlorinated- biphenyl-degrading Rhodococcus sp. strain RHA1.

Warren R, Hsiao W, Kudo H, Myhre M, Dosanjh M, Petrescu A J Bacteriol. 2004; 186(22):7783-95.

PMID: 15516593 PMC: 524921. DOI: 10.1128/JB.186.22.7783-7795.2004.

Characterization of the 101-kilobase-pair megaplasmid pKB1, isolated from the rubber-degrading bacterium Gordonia westfalica Kb1.

Broker D, Arenskotter M, Legatzki A, Nies D, Steinbuchel A J Bacteriol. 2003; 186(1):212-25.

PMID: 14679241 PMC: 303448. DOI: 10.1128/JB.186.1.212-225.2004.

DNA sequence and comparison of virulence plasmids from Rhodococcus equi ATCC 33701 and 103.

Takai S, Hines S, Sekizaki T, Nicholson V, Alperin D, Osaki M Infect Immun. 2000; 68(12):6840-7.

PMID: 11083803 PMC: 97788. DOI: 10.1128/IAI.68.12.6840-6847.2000.

References

Steinmetz M, Le Coq D, Aymerich S, Gay P . The DNA sequence of the gene for the secreted Bacillus subtilis enzyme levansucrase and its genetic control sites. Mol Gen Genet. 1985; 200(2):220-8. DOI: 10.1007/BF00425427. View

Li M, Squires C, Monticello D, Childs J . Genetic analysis of the dsz promoter and associated regulatory regions of Rhodococcus erythropolis IGTS8. J Bacteriol. 1996; 178(22):6409-18. PMC: 178525. DOI: 10.1128/jb.178.22.6409-6418.1996. View

Denome S, Oldfield C, Nash L, Young K . Characterization of the desulfurization genes from Rhodococcus sp. strain IGTS8. J Bacteriol. 1994; 176(21):6707-16. PMC: 197028. DOI: 10.1128/jb.176.21.6707-6716.1994. View

Jagura-Burdzy G, Khanim F, Smith C, Thomas C . Crosstalk between plasmid vegetative replication and conjugative transfer: repression of the trfA operon by trbA of broad host range plasmid RK2. Nucleic Acids Res. 1992; 20(15):3939-44. PMC: 334070. DOI: 10.1093/nar/20.15.3939. View

Jager W, Schafer A, Kalinowski J, Puhler A . Isolation of insertion elements from gram-positive Brevibacterium, Corynebacterium and Rhodococcus strains using the Bacillus subtilis sacB gene as a positive selection marker. FEMS Microbiol Lett. 1995; 126(1):1-6. DOI: 10.1111/j.1574-6968.1995.tb07381.x. View

Kulakov L, Larkin M, Kulakova A . Cryptic plasmid pKA22 isolated from the naphthalene degrading derivative of Rhodococcus rhodochrous NCIMB13064. Plasmid. 1997; 38(1):61-9. DOI: 10.1006/plas.1997.1293. View

Schagger H, von Jagow G . Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem. 1987; 166(2):368-79. DOI: 10.1016/0003-2697(87)90587-2. View

Piddington C, Kovacevich B, Rambosek J . Sequence and molecular characterization of a DNA region encoding the dibenzothiophene desulfurization operon of Rhodococcus sp. strain IGTS8. Appl Environ Microbiol. 1995; 61(2):468-75. PMC: 167304. DOI: 10.1128/aem.61.2.468-475.1995. View

Desomer J, Vereecke D, Crespi M, Van Montagu M . The plasmid-encoded chloramphenicol-resistance protein of Rhodococcus fascians is homologous to the transmembrane tetracycline efflux proteins. Mol Microbiol. 1992; 6(16):2377-85. DOI: 10.1111/j.1365-2958.1992.tb01412.x. View

10.

Zheng H, Prescott J . Development of a Rhodococcus equi-Escherichia coli plasmid shuttle vector. Plasmid. 1997; 38(3):180-7. DOI: 10.1006/plas.1997.1311. View

11.

Pelicic V, Reyrat J, Gicquel B . Expression of the Bacillus subtilis sacB gene confers sucrose sensitivity on mycobacteria. J Bacteriol. 1996; 178(4):1197-9. PMC: 177784. DOI: 10.1128/jb.178.4.1197-1199.1996. View

12.

Jager W, Schafer A, Puhler A, Labes G, Wohlleben W . Expression of the Bacillus subtilis sacB gene leads to sucrose sensitivity in the gram-positive bacterium Corynebacterium glutamicum but not in Streptomyces lividans. J Bacteriol. 1992; 174(16):5462-5. PMC: 206388. DOI: 10.1128/jb.174.16.5462-5465.1992. View

13.

Gay P, Le Coq D, Steinmetz M, Berkelman T, Kado C . Positive selection procedure for entrapment of insertion sequence elements in gram-negative bacteria. J Bacteriol. 1985; 164(2):918-21. PMC: 214340. DOI: 10.1128/jb.164.2.918-921.1985. View

14.

Pansegrau W, LANKA E, Barth P, Figurski D, Guiney D, Haas D . Complete nucleotide sequence of Birmingham IncP alpha plasmids. Compilation and comparative analysis. J Mol Biol. 1994; 239(5):623-63. DOI: 10.1006/jmbi.1994.1404. View

15.

Gallardo M, Ferrandez A, de Lorenzo V, Garcia J, Diaz E . Designing recombinant Pseudomonas strains to enhance biodesulfurization. J Bacteriol. 1997; 179(22):7156-60. PMC: 179658. DOI: 10.1128/jb.179.22.7156-7160.1997. View

16.

Scholz P, Haring V, Wittmann-Liebold B, Ashman K, Bagdasarian M, Scherzinger E . Complete nucleotide sequence and gene organization of the broad-host-range plasmid RSF1010. Gene. 1989; 75(2):271-88. DOI: 10.1016/0378-1119(89)90273-4. View

17.

Desomer J, DHaese P, Montagu M . Transformation of Rhodococcus fascians by High-Voltage Electroporation and Development of R. fascians Cloning Vectors. Appl Environ Microbiol. 1990; 56(9):2818-25. PMC: 184849. DOI: 10.1128/aem.56.9.2818-2825.1990. View

18.

Gray K, Pogrebinsky O, Mrachko G, Xi L, Monticello D, Squires C . Molecular mechanisms of biocatalytic desulfurization of fossil fuels. Nat Biotechnol. 1996; 14(13):1705-9. DOI: 10.1038/nbt1296-1705. View

19.

Qin M, Taniguchi H, Mizuguchi Y . Analysis of the replication region of a mycobacterial plasmid, pMSC262. J Bacteriol. 1994; 176(2):419-25. PMC: 205065. DOI: 10.1128/jb.176.2.419-425.1994. View

20.

Gavigan J, Ainsa J, Perez E, Otal I, Martin C . Isolation by genetic labeling of a new mycobacterial plasmid, pJAZ38, from Mycobacterium fortuitum. J Bacteriol. 1997; 179(13):4115-22. PMC: 179229. DOI: 10.1128/jb.179.13.4115-4122.1997. View