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Distribution of 2-kb miniplasmid pBMB2062 from Bacillus thuringiensis kurstaki YBT-1520 strain in Bacillus species

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Abstract

A multi-copy and small plasmid pBMB2062 from Bacillus thuringiensis kurstaki YBT-1520 strain was cloned and characterized and its distribution was analyzed using dot-blot analysis with the ORF1 fragment as a probe. Bacillus species of 84 serotypes were evaluated. The pBMB2062 plasmid was found to be present in commercial B. thuringiensis kurstaki (H3abc) and aizawai (H7) insecticides of various serotypes, and one Bacillus cereus UW85 strain (produced Zwittermicin fungicide and Cry toxin synergist). The sequences of 7 pBMB2062-like plasmids from randomly selected Bacillus species (positive signal in the dot-blot analysis) were highly conserved. Two open reading frames (ORFs), ORF1 and ORF2, were present in this plasmid. ORF1 was found to be necessary for plasmid replication, whereas ORF2 did not play a role in replication or stability. Based on its sequence homology, ORF2 was a putative solitary antitoxin. Furthermore, the copy number of the replicon of pBMB2062 was higher than those of ori1030 and ori44 based on the thermogenic data, and ori2062 could drive the stable replication of a recombinant plasmid (11 kb total size) in B. thuringiensis.

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References

  • Andrup L, Jensen GB, Wilcks A, Smidt L, Hoflack L, Mahillon J (2003) The patchwork nature of rolling-circle plasmids: comparison of six plasmids from two distinct Bacillus thuringiensis serotypes. Plasmid 49:205–232

    Article  PubMed  CAS  Google Scholar 

  • Arantes O, Lereclus D (1991) Construction of cloning vectors for Bacillus thuringiensis. Gene 108:115–119

    Article  PubMed  CAS  Google Scholar 

  • Baum JA, Gilbert MP (1991) Characterization and comparative sequence analysis of replication origins from three large Bacillus thuringiensis plasmids. J Bacteriol 173:5280–5289

    PubMed  CAS  Google Scholar 

  • Berry C, O’Neil S, Ben-Dov E, Jones AF, Murphy L, Quail MA, Holden MT, Harris D, Zaritsky A, Parkhill J (2002) Complete sequence and organization of pBtoxis, the toxin-coding plasmid of Bacillus thuringiensis subsp. israelensis. Appl Environ Microbiol 68:5082–5095

    Article  PubMed  CAS  Google Scholar 

  • Duan C, Tong Y, Huang Y, Wang X, Xiong X, Wen B (2011) Complete genome sequence of Rickettsia heilongjiangensis, an emerging tick-transmitted human pathogen. J Bacteriol 193:5564–5565

    Article  PubMed  CAS  Google Scholar 

  • Gerdes K (2000) Toxin–antitoxin modules may regulate synthesis of macromolecules during nutritional stress. J Bacteriol 182:561–572

    Article  PubMed  CAS  Google Scholar 

  • Gomis-Ruth FX, Sola M, Acebo P, Parraga A, Guasch A, Eritja R, Gonzalez A, Espinosa M, del Solar G, Coll M (1998) The structure of plasmid-encoded transcriptional repressor CopG unliganded and bound to its operator. EMBO J 17:7404–7415

    Article  PubMed  CAS  Google Scholar 

  • Grady R, Hayes F (2003) Axe-Txe, a broad-spectrum proteic toxin-antitoxin system specified by a multidrug-resistant, clinical isolate of Enterococcus faecium. Mol Microbiol 47:1419–1432

    Article  PubMed  CAS  Google Scholar 

  • Guerout-Fleury AM, Shazand K, Frandsen N, Stragier P (1985) Antibiotic-resistance cassettes for Bacillus subtilis. Gene 167:335–336

    Article  Google Scholar 

  • Handelsman J, Raffel S, Mester EH, Wunderlich L, Grau CR (1990) Biological control of damping-off of alfalfa seedlings with Bacillus cereus UW85. Appl Environ Microbiol 56:713–718

    PubMed  CAS  Google Scholar 

  • Khan SA (2003) DNA-protein interactions during the initiation and termination of plasmid pT181 rolling-circle replication. Prog Nucleic Acid Res Mol Biol 75:113–137

    Article  PubMed  CAS  Google Scholar 

  • Leonhardt H, Alonso JC (1991) Parameters affecting plasmid stability in Bacillus subtilis. Gene 103:107–111

    Article  PubMed  CAS  Google Scholar 

  • Lereclus D, Guo S, Sanchis V, Lecadet MM (1988) Characterization of two Bacillus thuringiensis plasmids whose replication is thermosensitive in B. subtilis. FEMS Microbiol Lett 49:417–422

    Article  CAS  Google Scholar 

  • Li L, Yu Z (1999) The transform and expressed property of Bacillus thuringiensis non-plasmid mutanant strain BMB171. Chin J Appl Environ Microbiol 5:395–399

    Google Scholar 

  • Lin XY, Liu Y, Liu P, Qu SS, Yu Z (2004) Microcalorimetric investigation on the growth model and the protein yield of Bacillus thuringiensis. J Biochem Bioph Meth 59:267–274

    Article  CAS  Google Scholar 

  • Lin XY, Liu Y, Sun M, Liu P, Zhang JC, Li L, Qu SS, Yu ZN (2005) Microcalorimetric investigation of the growth of the engineering Bacillus thuringiensis with different plasmid numbers and various promoters. J Therm Anal Calorim 79:29–32

    Article  Google Scholar 

  • Liu C, Bao QY, Song FP, Sun M, Huang DF, Liu GM, Yu ZN (2007) Complete nucleotide sequence of pBMB67, a 67-kb plasmid from Bacillus thuringiensis strain YBT-1520. Plasmid 57:44–54

    Article  CAS  Google Scholar 

  • Loeza-Lara PD, Benintende G, Cozzi J, Ochoa-Zarzosa A, Baizabal-Aguirre VM, Valdez-Alarcón JJ, López-Meza JE (2005) The plasmid pBMBt1 from Bacillus thuringiensis subsp. darmstadiensis (INTA Mo14-4) replicates by the rolling-circle mechanism and encodes a novel insecticidal crystal protein-like gene. Plasmid 54(3):229–240

    Article  PubMed  CAS  Google Scholar 

  • Marin R, Tanguay RM, Valero J, Letarte R, Bellemare G (1992) Isolation and sequence of a 2-kbp miniplasmid from Bacillus thuringiensis var. kurstaki HD-3a3b: relationship with miniplasmids of other B. thuringiensis strains. FEMS Microbiol Lett 73:263–269

    Article  PubMed  CAS  Google Scholar 

  • McDowell DG, Mann NH (1991) Characterization and sequence analysis of a small plasmid from Bacillus thuringiensis var. kurstaki strain HD1-DIPEL. Plasmid 25:113–120

    Article  PubMed  CAS  Google Scholar 

  • Rasko DA, Altherr MR, Han CS, Ravel J (2005) Genomics of the Bacillus cereus group of organisms. FEMS Microbiol Rev 29:303–329

    PubMed  CAS  Google Scholar 

  • Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor

    Google Scholar 

  • Sanchis V, Agaisse H, Chaufaux J, Lereclus D (1997) A recombinase-mediated system for elimination of antibiotic resistance gene markers from genetically engineered Bacillus thuringiensis strains. Appl Environ Mirobiol 63:779–784

    CAS  Google Scholar 

  • Sevin EW, Barloy-Hubler F (2007) RASTA-Bacteria: a web-based tool for identifying toxin-antitoxin loci in prokaryotes. Genome Biol 8:R155

    Article  PubMed  Google Scholar 

  • Shao Z, Yu Z (2004) Enhanced expression of insecticidal crystal proteins in wild Bacillus thuringiensis strains by a heterogeneous protein P20. Curr Microbiol 48:321–326

    Article  PubMed  CAS  Google Scholar 

  • Schnepf E, Crickmore N, Van Rie J, Lereclus D, Baum J, Feitelson J, Zeigler DR, Dean DH (1998) Bacillus thuringiensis and its pesticidal crystal proteins. Microbiol Mol Biol Rev 62:775–806

    PubMed  CAS  Google Scholar 

  • Sun M, Liu Z, Yu Z (2000a) Characterization of the insecticidal crystal protein genes of Bacillus thuringiensis YBT-1520. Wei Sheng Wu Xue Bao 40:365–371

    PubMed  CAS  Google Scholar 

  • Sun M, Wei F, Liu Z, Yu Z (2000b) Cloning of the pBMB2062 from B. thuringiensis and the construction of genetic stable vector. Acta Genet Sin 27:932–938

    PubMed  CAS  Google Scholar 

  • Sylvain E, Michel G, Didier L, Vincent S (2004) An extracytoplasmic-function sigma factor is involved in a pathway controlling β-exotoxin I production in Bacillus thuringiensis subsp. thuringiensis strain 407–1. J Bacteriol 186:3108–3116

    Article  Google Scholar 

  • Tang M, Bideshi DK, Park HW, Federici BA (2006) Minireplicon from pBtoxis of Bacillus thuringiensis subsp. israelensis. Appl Environ Microbiol 72:6948–6954

    Article  PubMed  CAS  Google Scholar 

  • Viret JF, Bravo A, Alonso JC (1991) Recombination-dependant concatemeric plasmid replication. Microbiol Rev 55:675–683

    PubMed  CAS  Google Scholar 

  • Wilcks A, Smidt L, Okstad OA, Kolsto AB, Mahillon J, Andrup L (1999) Replication mechanism and sequence analysis of the replicon of pAW63, a conjugative plasmid from Bacillus thuringiensis. J Bacteriol 181:3193–3200

    PubMed  CAS  Google Scholar 

  • Zhang Q, Sun M, Xu Z, Yu Z (2007) Cloning and characterization of pBMB9741, a native plasmid of Bacillus thuringiensis subsp. kurstaki strain YBT-1520. Curr Microbiol 55:302–307

    Article  PubMed  CAS  Google Scholar 

  • Zhong CY, Peng DH, Ye WX, Chai LJ, Qi JL, Yu ZN, Ruan LF, Sun M (2011) Determination of plasmid copy number reveals the total plasmid DNA amount is greater than the chromosomal DNA amount in Bacillus thuringiensis YBT-1520. PLoS One 6:e16025

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We thank Professor Jacques Mahillon (Catholic University of Louvain) for his personal communication on the TA system. We thank Mr. Yu Yao (Shanghai Jiaotong University) for his valuable help on article writing. We thank Dan Zeigler (Bacillus Genetics Stock Centers) and Jean-François Charles (Pasteur Institute) for providing the B. thuringiensis strains. This work was supported by grants from the National High Technology Research and Development Program (863) of China (2011AA10A203), China 948 Program of Ministry of Agriculture (2011-G25), the National Basic Research Program (973) of China (2009CB118902) and the National Natural Science Foundation of China (31170047 and 31000020).

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  1. State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, 430070, Wuhan, People’s Republic of China

    Xiao-Jin Liu, Li-Fang Ruan, Xiao-Yan Lin, Chang-Ming Zhao, Chun-Ying Zhong & Ming Sun

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  1. Xiao-Jin Liu
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  2. Li-Fang Ruan
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Correspondence to Ming Sun.

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Liu, XJ., Ruan, LF., Lin, XY. et al. Distribution of 2-kb miniplasmid pBMB2062 from Bacillus thuringiensis kurstaki YBT-1520 strain in Bacillus species. Ann Microbiol 63, 1639–1644 (2013). https://doi.org/10.1007/s13213-013-0627-8

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