A vector with the downstream box of the initiation codon can highly enhance protein expression in Escherichia coli.
An expression vector, pET-DB, with a perfectly matching downstream box of the initiation codon has been constructed on the basis of the pET system. Any gene of interest can then be inserted into the vector. Four genes were used to test the expression efficiency of the vector. The results show that the vector pET-DB can further increase protein expression level at least up to 35-70% as compared with the initial T7 expression system, indicating that the downstream box can enhance protein expression in Escherichia coli.
Zhang
Xiaolan
X
National Laboratory of Biomacromolecules, Institute of Biophysics, Academia Sinica, Beijing 100101, P.R. China.
Guo
Peng
P
Jing
Guozhong
G
D000263:Adenylate Kinase / Q000235:genetics / Q000378:metabolism; D000818:Animals; D051017:Apoptosis Regulatory Proteins; D001483:Base Sequence; D002645:Chickens; D003013:Clostridium / Q000235:genetics / Q000378:metabolism; D018387:Codon, Initiator / Q000235:genetics; D004926:Escherichia coli / Q000145:classification / Q000235:genetics* / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics; D017930:Genes, Reporter; D005822:Genetic Vectors; D006801:Humans; D008836:Micrococcal Nuclease / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000201:enzymology; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism; D010442:Peptide Chain Initiation, Translational / Q000235:genetics*; D014176:Protein Biosynthesis; D011506:Proteins / Q000235:genetics*; D012329:RNA, Bacterial / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D012358:RNA, Transfer, Met; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
D051017:Apoptosis Regulatory Proteins; D018387:Codon, Initiator; D009363:Neoplasm Proteins; C117283:PDCD5 protein, human; D011506:Proteins; D012329:RNA, Bacterial; D012333:RNA, Messenger; D012358:RNA, Transfer, Met; D011993:Recombinant Fusion Proteins; D000263:Adenylate Kinase; D008836:Micrococcal Nuclease