概要
- 理化学研究所 播磨研究所 放射光科学総合研究センター/大阪大学 倉光成紀先生より寄託していただいた、”高度好熱菌 (Thermus thermophilus HB8) 丸ごと一匹プロジェクト” で構築された、組換えタンパク質を大腸菌で発現するためのプラスミドならびに遺伝子破壊用プラスミドです。
Expression plasmid of Thermus thermophilus HB8 proteins in E. coli
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 |
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| Related information |
Please visit Informaion site  for articles published by using this materials, references and tips.
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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
| 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 |
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| 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.
提供申込方法
生物遺伝資源提供依頼書類の入手と記入
次の書式に記入してください:
- 提供依頼書
- 提供を希望される「クローンの番号 (例：TEx01A01, TDs01A01)を記入
- カタログNo. (RDB)欄の記入は不要
書式A （遺伝子材料提供依頼書） [Word] [依頼書の記入例] 1部 - 生物遺伝資源提供同意書 (MTA)
- リソース名は「Thermus thermophilus遺伝子発現プラスミド（別添参照）」を記入
- 固有記号欄の記入は不要
- リソースの利用条件欄には次の1点を記入
「利用者は、研究成果の公表にあたって寄託者が指定する文献を引用する (Yokoyama et al., 2000, Nature Struct. Biol. 7, 943-945)。」 - 利用目的により、非営利学術目的用あるいは営利目的用を選択してください。
書式C (生物遺伝資源提供同意書, MTA) 非営利学術目的用 [Word]
書式C (生物遺伝資源提供同意書, MTA) 営利目的用 [Word]2部
書類の送付とお問い合わせ
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提供手数料：（1本あたり)
| 非営利学術目的利用 | 9,460円 |
| 営利目的利用 | 18,920円 |
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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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2023.06.11
