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.
Thermus thermophilusHost–vector system Colour screening β-Galactosidase S-gal
Multiple cloning sites
Polymerase chain reaction
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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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