Tandem repeating of the expression cartridge--a novel strategy to enhance the expression efficiency of cloned gene in Escherichia coli.
A novel strategy to enhance the expression efficiency of cloned target gene in Escherichia coli was developed. The whole expression cartridge, consisting of promoter, SD sequence, target gene and transcription terminator, was tandem repeatedly engineered into a expression plasmid. Consequently, the copy number of specific gene was increased substantially, leading to the improvement of expression efficiency. Using this approach, a recombinant plasmid, designed as pLYD, was constructed and transformated into the Escherichia coli strain DH5 alpha. Upon induction, the desired protein was synthesized in a considerable level and accumulated up to 63% of the total cell proteins. The present study revealed that tandem repeating of expression cartridge provided a convenient means to improve expression level efficiently.
Lin
Y
Y
Institute of Basic Medical Sciences, CAMS, Beijing.
Lu
S
S
D004274:DNA, Recombinant; D004926:Escherichia coli / Q000235:genetics*; D005818:Genetic Engineering; D010957:Plasmids; D012091:Repetitive Sequences, Nucleic Acid; D014170:Transformation, Genetic
D004274:DNA, Recombinant