Polylinker-encoded peptides can confer toxicity to recombinant proteins produced in Escherichia coli.
Three DNA segments encoding fragments of the p60 capsid protein of rabbit haemorrhagic disease virus (RHDV) have been cloned and expressed in Escherichia coli. The cDNAs were placed under the control of the T7 promoter in a pET-derived expression vector designed to produce C-terminal histidine tail fusions. By using different cloning strategies, cell toxicity exhibited by some of the expressed products was dramatically affected, coincident with minor modifications in the carboxy terminus sequences of the recombinant proteins. In particular, the presence of a nonhydrophobic peptide encoded by polylinker sequences promotes cell death and a reduced yield after induction of gene expression, whereas a histidine tail has no detectable effect. These data point out the critical role that needless peptide tails, accidentally introduced into recombinant proteins by nonrelevant DNA stretches, can have on protein expression and final yield of a production process.
Viaplana
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E
Institut de Biologia Fonamental, Universitat Autònoma de Barcelona, Spain.
Villaverde
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A
D000595:Amino Acid Sequence; D000818:Animals; D002213:Capsid / Q000378:metabolism; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000235:genetics; D004926:Escherichia coli / Q000254:growth & development / Q000378:metabolism*; D015870:Gene Expression; D005814:Genes, Viral; D017929:Hemorrhagic Disease Virus, Rabbit / Q000235:genetics*; D008969:Molecular Sequence Data; D010957:Plasmids; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000633:toxicity; D014764:Viral Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000633:toxicity*
D004279:DNA, Viral; D011994:Recombinant Proteins; D014764:Viral Proteins