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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

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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.

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Abbreviations

bp:

Base pair

MCS:

Multiple cloning sites

PCR:

Polymerase chain reaction

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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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Authors and Affiliations

  1. Biomedical Research Institute, Health Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-8-31 Midorigaoka, Ikeda, Osaka, 563-8577, Japan

    Atsushi Fujita

  2. Department of Cell Biology, Fukuoka University School of Medicine, 7-45-1 Nanakuma, Jonan-ku, Fukuoka, 814-0180, Japan

    Yoshio Misumi

Authors
  1. Atsushi Fujita
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  2. Yoshio Misumi
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Correspondence to Atsushi Fujita.

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Communicated by L. Huang.

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Fujita, A., Misumi, Y. 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. Extremophiles 21, 1111–1117 (2017). https://doi.org/10.1007/s00792-017-0961-z

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  • DOI: https://doi.org/10.1007/s00792-017-0961-z

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