1 A functional recT gene for recombineering of Clostridium

2
3 Hongjun Dong1, Wenwen Tao1,2, Fuyu Gong1,2, Yin Li1, Yanping Zhang1*
4 1CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of
5 Microbiology, Chinese Academy of Sciences, Beijing, China.
6 2University of Chinese Academy of Sciences, Beijing, China.
7 *To whom correspondence should be addressed. Yanping Zhang, Institute of
8 Microbiology, Chinese Academy of Sciences, No. 1 West Beichen Road, Chaoyang District,
9 Beijing 100101, China. E-mail: zhangyp@im.ac.cn. Tel: +86-10-64807451. Fax:
10 +86-10-64807451.
11 Abstract
12 Recombineering is an efficient genetic manipulation method employing the mechanism of
13 phagenic RecT-mediated homologous recombination. To develop a recombineering method for
14 Clostridium, a putative recT gene (CPF0939) from C. perfringens genome was functionally
15 verified in a clostridial host C. acetobutylicum. We show that a short synthetic oligonucleotide can
16 be introduced into the target site for specific point mutation. This functional recT gene would
17 therefore contribute to development of recombineering tools for Clostridium.