DNA Vectors

pUT mini-Tn5 Vectors

pUT mini-Tn5 system

The mini-Tn5 are mobile elements able to transpose to the chromosome from a delivery plasmid, by means of the transposase activity encoded by the tnp* gene, which is present in cis in the plasmid but external to the mini-Tn5 element. pUT mini-Tn5 vectors can be used to insert any DNA fragment in the bacterial chromosome, by cloning it within the mini-Tn5 region. Due to the loss of the tnp* gene after insertion, minitransposons are stably integrated into the chromosome and inherited. The different pUTmini-Tn5 vectors can then be used for repeated insertion events and therefore multiple insertions in the same strain are only limited by the availability of distinct selection markers.

Advantages:

  • Easy and stable insertion of heterologous DNA in the chromosome of the strain of interest, after few simple genetic manipulations.
  • Wide range of inserted fragment sizes.
  • Possibility of multiple insertions in the same strain.
  • Easy modification of the cloning vector.
  • Use of cleavage sites of the rare cutters NotI and SfiI.
  • Wide range of Gram negative recipient strains.

Some of the most common applications of pUTmini-Tn5 vectors are:

  • Insertional mutagenesis.
  • Promoter probing.
  • Generation of conditional phenotypes and non-polar mutations.
  • Analysis of transcriptional terminators.
  • Environmental applications: tagging with marker genes for monitoring of released microorganisms and/or transcriptional regulation in the environment.

Bibliography:

pUT mini-Tn5 kits

pUT mini-Tn5 Cm kit

  • pUT mini-Tn5 Cm, 2 μ
  • coli DH5αlpir, stab.
  • coli S17-1 lpir, stab.
  • coli DH5α(pRK2013), stab.
  • TSS solution, 1.5ml.

pUT mini-Tn5 Km kit

  • pUT mini-Tn5 Km, 2 μ
  • E.coli DH5a lpir, stab.
  • E.coli S17-1 lpir, stab.
  • E.coli DH5a(pRK2073), stab.
  • TSS solution, 1.5ml.

pUT mini-Tn5 Sm/Sp kit

  • pUT mini-Tn5 Sm/Sp, 2 μ
  • E.coli DH5alpir, stab.
  • E.coli S17-1 lpir, stab.
  • E.coli DH5a(pRK2013), stab.
  • TSS solution, 1.5ml.

pUT mini-Tn5 Tc kit

  • pUT mini-Tn5 Tc, 2 μ
  • E.coli DH5alpir, stab.
  • E.coli S17-1 lpir, stab.
  • E.coli DH5a(pRK2013), stab.
  • TSS solution, 1.5ml.

PANC-1 mini-Tn5 package

  • pUT mini-Tn5 Km, 2 μ
  • pUT mini-Tn5 Cm, 2 μ
  • pUT mini-Tn5 Sm/Sp, 2 μ
  • pUT mini-Tn5 Tc, 2 μ

PANC-1plus mini-Tn5 package

  • pUT mini-Tn5 Km, 2 μ
  • pUT mini-Tn5 Cm, 2 μ
  • pUT mini-Tn5 Sm/Sp, 2 μ
  • pUT mini-Tn5 Tc, 2 μ
  • E.coli DH5alpir, stab.
  • E.coli S17-1 lpir, stab.
  • E.coli DH5a(pRK2013), stab.
  • E.coli DH5a(pRK2073), stab.
  • TSS solution, 1.5ml.

pUT mini-Tn5Tel kit

  • pUT mini-Tn5 Tel, 2 μ
  • Potassium Tellurite, 100mg.
  • E.coli DH5alpir, stab.
  • E.coli S17-1 lpir, stab.
  • E.coli DH5a(pRK2013), stab.
  • TSS solution, 1.5ml.

Related products

Related Strains

BS-3233       E.coli DH5alpir     Stab
BS-3234       E.coli S17-1 lpir     Stab
BS-3236       E.coli DH5a(pRK2013)     Stab
BS-3263       E.coli DH5a(pRK2073)     Stab
KT-4694      E.coli DH5a Competent Cells Set     12 vials of 0.2 ml and pUC18Not

Related Reagents

RS-3215 TSS solution – 20 reactions     1.5 ml
RS-3216 TSS solution – 100 reactions     1.5 ml x 5
RS-3445 Potassium Tellurite     100mg
RS-3222 Potassium Tellurite     1g
RS-3311 Potassium Tellurite     5g
RS-3217 Ampicillin     5 g
RS-3305 Ampicillin     25g
RS-3306 Ampicillin     100g
RS-3218 Chloramphenicol     5 g
RS-3307 Chloramphenicol     100g
RS-3315 Erythromycin     5g
RS-3327 Erythromycin     100g
RS-3328 Gentamycin     1g
RS-3329 Gentamycin     5g
RS-3219 Kanamycin     5 g
RS-3221 Streptomycin     5 g
RS-3310 Streptomycin     25g
RS-3220 Tetracycline     5 g
RS-3308 Tetracycline     25g
RS-3309 Tetracycline     100g
RS-3222 Potassium Tellurite     1 g
RS-3311 Potassium Tellurite     5g