Construction of a novel shuttle vector for use in Brevibacterium flavum, an industrial amino acid producer.
Brevibacterium flavum has been developed to produce amino acids l-valine, l-lysine and l-threonine. However, there are not enough vectors available for the research on metabolic engineering in Brevibacterium flavum. Here we have constructed a shuttle expression vector pDXW-8 between Escherichia coli and B. flavum. The vector harbors an origin oriE for replicating in E. coli, a second origin oriC for replicating in B. flavum, a large multiple cloning site including 11 single restriction enzyme sites and suitable for cloning multiple genes or large DNA fragments, a tac promoter and a lacI(PF104) fragment which tightly controls the tac promoter. The applicability of pDXW-8 was confirmed by the expression of the vhb gene in B. flavum. The vector pDXW-8 will be very useful for research on metabolic engineering in corynebacteria.
Xu
Daqing
D
State Key Laboratory of Food Science and Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi 214122, China.
Tan
Yanzhen
Y
Huan
Xiaojing
X
Hu
Xiaoqing
X
Wang
Xiaoyuan
X
D000596:Amino Acids / Q000378:metabolism*; D040142:Brevibacterium flavum / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D007218:Industrial Microbiology
D000596:Amino Acids