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Thermus thermophilus Expression & Disruption plasmid

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Thermus thermophilus HB8 Expression and Disruption plasmids were constructed by “Whole-Cell Project of a Model Organism, Thermus thermophilus HB8” and deposited by Dr. Seiki Kuramitsu.

Expression plasmid of Thermus thermophilus HB8 proteins in E. coli

Data Sheet data sheet

Resource name Thermus thermophilus HB8 expression plasmid
Vector pET-11a (Ampr), pET-11b (Ampr) or pET-3a (Ampr)
Insert Genomic DNA of Thermus thermophilus strain HB8
Host Plasmids were amplified using DH5alpha, E. coli
Individual clone data Available at Thermus thermophilus gene plasmid
Reference
  • Yokoyama, S., Hirota, H., Kigawa, T., Yabuki, T., Shirouzu, M., Terada, T., Ito, Y., Matsuno, Y. Kuroda, Y., Nishimura, Y., Kyogoku, Y., Miki, K., Masui, R., Kuramitsu, S. (2000) Structual genomics projects in Japan. Nature Struct. Biol. 7, 943-945.
  • Structural-Biological Whole Cell Project
Related information Please visit Informaion site for articles published by using this materials, references and tips.

Disruption plasmid of Thermus Thermophilus HB8 gene

The gene disruptant of T. thermophils HB8 can be easily prepared by adding this plasmid into the culture medium. The target gene in this plasmid was replaced by the thermostable kanamycin resistant gene from Staphylococcus aureus. The length of the homologous region outside of the target gene is about 500 bp (only 10 bp of the target geneare left in both sides) .

Data Sheet data sheet

Resource name Thermus thermophilus HB8 Disruption plasmid
Vector pGEM derivative (Ampr)
Insert Genomic DNA of Thermus thermophilus strain HB8, thermostable kanamycin resistant gene from Staphylococcus aureus
Host Plasmids were amplified using E. coli host strain for unstable inserts (ex. Stbl2)
Individual clone data Available at Thermus thermophilus gene plasmid
Sequencing Primer
  • Vector toward insert
    T7long: cgccaagctctaatacgactcactataggg
    SP6: atttaggtgacactatag
  • Km toward insert
    7723km F：aatttctggaatggggttca
    7723km R：gattgcgatgctgattcgt
Related information Please visit Informaion site for articles published by using this materials, references and tips.

Disruption method

Directed evolution of thermostable kanamycin-resistance gene: a convenient selection marker for Thermus thermophilus.

Hoseki, J., Yano, T., Koyama, Y., Kuramitsu, S., Kagamiyama, H.
J. Biochem. 126 (5): 951-956 (1999). PubMed PMID 10544290.

Catalog no. Name of resource Description
RDB03436 pUC18-HTK (with promoter) Expression vector of Thermus highly thermostable kanamycin nucleotidyltransferase (HTK) gene

Depositor: Dr. Jun Hoseki

Related article

  • Molecular mechanisms of the whole DNA repair system: a comparison of bacterial and eukaryotic systems.
    Shimada, A., Inoue, M., Iino, H., Wakamatsu, T., Fukui, K., Nakagawa, N., Masui, R., Kuramitsu, S. (2010)
    J. Nucleic Acids 2010: 179594
  • Increased Rigidity of Domain Structures Enhances the Stability of a Mutant Enzyme Created by Directed Evolution
    Hoseki, J., Okamoto, A., Takada, N., Suenaga, A., Funatsugi, N., Konagaya, A., Taiji, M., Yano, T., Kuramitsu, S. and Kagamiyama, H. (2003)
    Biochemistry 42, 14469-14475.
  • Disruption of Thermus thermophilus Genes by Homologous Recombination Using A Thermostable Kanamycin-Resistant Marker
    Hashimoto, Y., Yano, T., Kuramitsu, S. and Kagamiyama, H. (2001)
    FEBS Lett. 506, 231-234.
  • Directed Evolution of Thermostable Kanamycin-Resistant Gene Products in an Extremely Thermophilic Bacterium, Thermus thermophilus
    Hoseki, J., Yano, T., Koyama, Y., Kuramitsu, S. and Kagamiyama, H. (1999)
    J. Biochem. 126, 951-956.

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Distribution fee per clone (as of April 1st, 2023):
JPY 9,460 (For use in research for not-for-profit academic purpose).
JPY 18,920 (For use in research for-profit-research purpose).
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TIPS

  • Strain Thermus thermophilus HB8 (JCM 10941T) of BRC-JCM
  • Genomic DNA (cat# JGD05989) of Thermus thermophilus HB8 (JCM 10941T)

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(GRP0051e 2012.10.22 T.M.)

2023.06.08



 

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