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

被引：1
作者：
Fujita, Atsushi [1 ]
Misumi, Yoshio [2 ]
机构：
[1] Natl Inst Adv Ind Sci & Technol, Hlth Res Inst, Biomed Res Inst, 1-8-31 Midorigaoka, Ikeda, Osaka 5638577, Japan
[2] Fukuoka Univ, Sch Med, Dept Cell Biol, Jonan Ku, 7-45-1 Nanakuma, Fukuoka 8140180, Japan
来源：
EXTREMOPHILES | 2017年 / 21卷 / 06期
关键词：
Thermus thermophilus; Host-vector system; Colour screening; beta-Galactosidase; S-gal; ESCHERICHIA-COLI; EXTREME THERMOPHILE; MARKER; TRANSFORMATION; KANAMYCIN;
D O I：
10.1007/s00792-017-0961-z
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
We constructed a new Thermus thermophilus cloning vector which enables the colour selection of cloned DNA inserts in the T. thermophilus HB27 host strain (beta-gal(-)) on growth plates containing 3,4-cyclohexenoesculetin beta-d-galactopyranoside (S-gal) in the medium. This vector harbors a modified beta-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.
引用
收藏
页码：1111 / 1117
页数：7
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