Establishment of a Genetic Transformation System and Its Application in Thermoanaerobacter tengcongensis

Bo Liu^{a, b},
Chuan Wang^a,
Haihua Yang^a,
Huarong Tan^{a
, ,}

a.
State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China
b.
Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China

More Information

Corresponding author: E-mail address: tanhr@im.ac.cn (Huarong Tan)

摘要

- /
Abstract: The whole-genome sequence of Thermoanaerobacter tengcongensis, an anaerobic thermophilic bacterium isolated from the Tengchong hot spring in China, was completed in 2002. However, in vivo studies on the genes of this strain have been hindered in the absence of genetic manipulation system. In order to establish such a system, the plasmid pBOL01 containing the replication origin of the T. tengcongensis chromosome and a kanamycin resistance cassette, in which kanamycin resistance gene expression was controlled by the tte1482 promoter from T. tengcongensis, was constructed and introduced into T. tengcongensis via electroporation. Subsequently, the high transformation efficiency occurred when using freshly cultured T. tengcongensis cells without electroporation treatment, suggesting that T. tengcongensis is naturally competent under appropriate growth stage. A genetic transformation system for this strain was then established based on these important components, and this system was proved to be available for studying physiological characters ofT. tengcongensis in vivo by means of hisG gene disruption and complementation.
- Genetic transformation system /
- Gene disruption and complementation

HTML全文

参考文献(24)

[1]	Bao, Q.Y., Tian, Y.Q., Li, W. et al. Genome Res., 12 (2002),pp. 689-700
[2]	Bradley, M.D., Beach, M.B., de Koning, A.P.J. et al. Microbiology, 153 (2007),pp. 2922-2940
[3]	Chen, Q., You, D.L., Hua, M.H. et al. Protein Expr. Purif., 32 (2003),pp. 239-245
[4]	Georgieva, T.I., Ahring, B.K. Appl. Microbiol. Biotechnol., 77 (2007),pp. 61-68
[5]	Grado, M.D., Castan, P., Berenguer, J. Plasmid, 42 (1999),pp. 241-245
[6]	Havarstein, L.S., Coomaraswamy, G., Morrison, D.A. Proc. Natl. Acad. Sci. USA, 92 (1995),pp. 11140-11144
[7]	Kahramanoglou, C., Seshasayee, A.S., Prieto, A.I. et al. Nucleic Acids Res., 39 (2011),pp. 2073-2091
[8]	Klapatch, T.R., Guerinot, M.L., Lynd, L.R. J. Ind. Microbiol., 16 (1996),pp. 342-347
[9]	Liang, C.N., Fioroni, M., Ropero, F.R. et al. Directed evolution of a thermophilic endoglucanase (Cel5A) into highly active Cel5A variants with an expanded temperature profile J. Biotechnol., 154 (2011),pp. 46-53
[10]	Mai, V., Lorenz, W.W., Wiegel, J. FEMS Microbiol. Lett., 148 (1997),pp. 163-167
[11]	Meng, B., Qian, Z., Wei, F. et al. Proteomics, 9 (2009),pp. 3189-3200
[12]	Oberholzer, A.E., Bumann, M., Schneider, P. et al. J. Mol. Biol., 346 (2005),pp. 521-532
[13]	Palmen, R., Buijsman, P., Hellingwerf, K.J. Arch. Microbiol., 162 (1994),pp. 344-351
[14]	Pei, H.D., Liu, J.F., Li, J. et al. Nucleic Acids Res., 35 (2007),pp. 3087-3099
[15]	Peng, H., Fu, B.L., Mao, Z.G. et al. Biotechnol. Lett., 28 (2006),pp. 1913-1917
[16]	Revill, W.P., Bibb, M.J., Scheu, A.K. et al. J. Bacteriol., 183 (2001),pp. 3526-3530
[17]	Royter, M., Schmidt, M., Elend, C. et al. Extremophiles, 13 (2009),pp. 769-783
[18]	Shaw, A.J., Hogsett, D.A., Lynd, L.R. Appl. Environ. Microbiol., 76 (2010),pp. 4713-4719
[19]	Wang, J.Q., Xue, Y.F., Feng, X.L. et al. Proteomics, 4 (2004),pp. 136-150
[20]	Wang, J.Q., Zhao, C.F., Meng, B. et al. Proteomics, 7 (2007),pp. 1409-1419
[21]	Wolterink-van Loo, S., Levisson, M., Cabrieres, M.C. et al. Extremophiles, 12 (2008),pp. 461-469
[22]	Xue, Y.F., Xu, Y., Liu, Y. et al. Int. J. Syst. Evol. Microbiol., 51 (2001),pp. 1335-1341
[23]	Yao, W., Shi, L., Liang, D.C. Biochem. Biophys. Res. Commun., 361 (2007),pp. 1027-1032
[24]	Zhang, S.L., Liu, B., Yang, H.H. et al. Nucleic Acids Res., 38 (2010),pp. 3709-3720