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Thermus thermophilus Disruption plasmid


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


Thermus thermophilus HB8 Disruption plasmids were constructed by "Whole-Cell Project of a Model Organism, Thermus thermophilus HB8" (http://www.thermus.org/) using the thermostable kanamycin nucleotidyltransferase gene and deposited by Dr. Seiki Kuramitsu.

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) .

How to obtain Disruption clones


This page

Plasmid clones for disrupting Thermus thermophilus HB8 gene.
Clone(s) is shipped as DNA solution in TE buffer in individual tube.
Please amplify this plasmid clone using E. coli host strain (ex. Stbl2, Invitrogen) suitable for the cloning of unstable inserts such as direct repeats.

Outside database

Please find ID of individual clone(s) using the following outside database
http://www.tanpaku.org/ThermusDB/index.html
or this summary table.

Forms

Please specify ID (ex. TDs01A01) of individual clone(s).

Please indicate Terms and Conditions for Distribution in MTA as follows:

In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, the USER is expected to cite the following literature.
(Yokoyama et al., 2000, Nature Struct. Biol. 7, 943-945).

Please complete forms:

Order Form, FormA
Material Transfer Agreement, FormC
Order Form (FormA) [Open a new window!!]
Please complete the form with your shipping information including your account number of an international courier (FedEx. World Courier, TNT Express, DHL Global Forwarding and others).
NOTE: User registration on our system is required before download the Order Form for credit card (Visa / Master).
Material Transfer Agreement (Category I MTA) [Word]
For the use of our bioresource in research for not-for-profit academic purpose by a non-profit organization.

We ask a signature of the Authorized Representative of a recipient institution.
In the Section 2(a), please write your purpose of use of clone in 10 to 20 words.

(Please e-mail to dnabank.brc@riken.jp to obtain MTA in MS-Word format)
Material Transfer Agreement (Category II MTA) [Word]
For the use of our bioresource in research for the following cases:
 1. For research to be conducted by for-profit organizations.
 2. For collaborative research between for-profit organization and not-for-profit organization.
 3, For research by not-for- organization outsourced and sponsored by for-profit organization.
 4. For for-profit research by not-for-profit organization including R&D with the aim of patent acquisition.

Send forms:

The DNA Bank, RIKEN BioResource Research Center,
3-1-1 Koyadai, Tsukuba, Ibaraki 305-0074, Japan
E-mail: dnabank.brc@riken.jp
FAX: (+81)-29-836-9120

Please visit further information of distribution and fees.

Distribution fee per clone:
8,640 YEN (For use in research for not-for-profit academic purpose).
17,280 YEN (For use in research for-profit-research purpose).
(Shipping cost is not included.)

For details for fee and payment, please visit Information of Request for Distribution.
The distribution fees will be revised from October 1, 2019

Personal data protection policy

For protection of personal data at RIKEN BioResource Research Center, please reffer Personal data protection policy.

data sheet

Data Sheet

Vector pGEM derivative
thermostable kanamycin resistant gene from Staphylococcus aureus
Sequencing Primer (Vector toward insert) T7long: cgccaagctctaatacgactcactataggg SP6: atttaggtgacactatag
Sequencing Primer (Km toward insert) 7723km F:aatttctggaatggggttca 7723km R:gattgcgatgctgattcgt

Related information

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.
  • Please visit Informaion site for articles published by using this materials, references and tips.

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(GRP0035e, 2012.10.22 T.M.)

2019.08.12