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MuA Transposase

Domus MuA Transposase is a highly purified, native MuA enzyme. Domus offers MuA Transposase in ready-to-use concentration for in vitro – and in vivo applications.

Catalog Number Unit Size Usage  
D010101 2.2 µg (0.22 µg/µl) in vitro applications  Buy now
       
D010102 4 µg (0.4 µg/µl) in vivo applications  Buy now
       

In vitro Applications

  • In vitro insertion of Mu transposon into linear DNA or plasmids
  • In vitro insertion of Mu transposon into large genomic clones (e.g. BACs, YACs)

Benefits

  • Simple in vitro reaction, with only transposon DNA, target DNA, MuA Transposase and a simple reaction buffer (1,2)
  • Nearly random insertion profile (3,4)
  •  

Scheme for in vitro transposition integration.  A tetramer of MuA transposase and Mu transposon ends assemble into a stable transpososome. Under reaction conditions with Mg2+, the transpososome becomes activated and executes transposon integration into the target DNA.

In vivo Applications

  • Preparation of Mu transpososomes for efficient gene delivery following electroporation

Benefits

  • Efficient gene delivery for a variety of bacteria, and also for yeast and mammalian cells (5-10)
  • Can be used to generate exhaustive insertion mutant libraries for many types of micro-organisms, also to archaeal species (10)
  • Nearly random insertion profile (3,4)

Scheme for genomic integration. A tetramer of MuA transposase and Mu transposon ends assemble into stable transpososomes in vitro. Following electroporation, the transpososomes encounter Mg2+ ions in vivo and integrate transposon DNA into the chromosome.

 

References

1. Savilahti H, Rice PA, Mizuuchi K (1995) The phage Mu transpososome core: DNA requirements for assembly and function. EMBO J 14:4893-4903
2. Haapa S, Taira S, Heikkinen E, Savilahti H (1999) An efficient and accurate integration of mini-Mu transposons in vitro: a general methodology for functional genetic analysis and molecular biology applications. Nucleic Acids Res 27:2777-2784
3. Haapa-Paananen S, Rita H, Savilahti H (2002) DNA transposition of bacteriophage Mu. A quantitative analysis of target site selection in vitro. The Journal of Biological Chemistry 277:2843-2851
4. Mizuuchi M, Mizuuchi K (1993) Target site selection in transposition of phage Mu. Cold Spring Harb Symp Quant Biol 58:515-523
5. Lamberg A, Nieminen S, Qiao M, Savilahti H (2002) Efficient insertion mutagenesis strategy for bacterial genomes involving electroporation of in vitro-assembled DNA transposition complexes of bacteriophage Mu. Appl Environ Microbiol 68:705-712
6. Pajunen MI, Pulliainen AT, Finne J, Savilahti H (2005) Generation of transposon insertion mutant libraries for Gram-positive bacteria by electroporation of phage Mu DNA transposition complexes. Microbiology 151:1209-1218
7. Paatero AO, Turakainen H, Happonen LJ, Olsson C, Palomäki T, Pajunen MI, Meng X, Otonkoski T, Tuuri T, Berry C, Malani N, Frilander MJ, Bushman FD, Savilahti H (2008) Bacteriophage Mu integration in yeast and mammalian genomes. Nucleic Acids Res 36:e148
8. Tu Quoc PH, Genevaux P, Pajunen M, Savilahti H, Georgopoulos C, Schrenzel J, Kelley WL (2007) Isolation and characterization of biofilm formation-defective mutants of Staphylococcus aureus. Infect Immun 75:1079-1088
9. Wu Z, Xuanyuan Z, Li R, Jiang D, Li C, Xu H, Bai Y, Zhang X, Turakainen H, Saris PE, Savilahti H, Qiao M (2009) Mu transposition complex mutagenesis in Lactococcus lactis-identification of genes affecting nisin production. J Appl Microbiol 106:41-8
10. Kiljunen S, Pajunen MI, Dilks K, Storf S, Pohlschroder M, Savilahti H (2014) Generation of comprehensive transposon insertion mutant library for the model archaeon, Haloferax volcanii , and its use for gene discovery. BMC Biol 12:103

 For Research Use Only. Not for use in diagnostic procedures.