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Extremophiles

, Volume 21, Issue 6, pp 1111–1117 | Cite as

Development of a new host–vector system for colour selection of cloned DNA inserts using a newly designed β-galactosidase gene containing multiple cloning sites in Thermus thermophilus HB27

  • Atsushi Fujita
  • Yoshio Misumi
Method Paper

Abstract

We constructed a new Thermus thermophilus cloning vector which enables the colour selection of cloned DNA inserts in the T. thermophilus HB27 host strain (β-gal) on growth plates containing 3,4-cyclohexenoesculetin β-d-galactopyranoside (S-gal) in the medium. This vector harbors a modified β-galactosidase gene (TTP0042 of T. thermophilus HB27) with 12 unique restriction enzyme sites (Acc65I, AvrII, BlpI, BssHII, EcoRI, EcoRV, HindIII, NruI, SalI, SpeI, SphI and XbaI) as multiple cloning sites under the control of the T. thermophilus slpA promoter. This host–vector system facilitates cloning procedures in T. thermophilus HB27.

Keywords

Thermus thermophilus Host–vector system Colour screening β-Galactosidase S-gal 

Abbreviations

bp

Base pair

MCS

Multiple cloning sites

PCR

Polymerase chain reaction

Notes

Acknowledgements

We are grateful to Mr. Jiro Hasegawa for technical assistance, with illustrations. We would like to thank Editage (http://www.editage.jp) for English language editing.

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Copyright information

© Springer Japan KK 2017

Authors and Affiliations

  1. 1.Biomedical Research Institute, Health Research InstituteNational Institute of Advanced Industrial Science and Technology (AIST)IkedaJapan
  2. 2.Department of Cell BiologyFukuoka University School of MedicineFukuokaJapan

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