Low level expression in Escherichia coli of a fungal gene under the control of strong promoters.
We have chosen the Isopenicillin N synthetase (IPNS) gene from Cephalosporium acremonium to study its expression in Escherichia coli due to its peculiar DNA sequence [1]. Significant levels of the protein could not be detected when the IPNS gene was placed under the control of strong promoters. Only when the construction was cloned in a runaway plasmid, significant levels of the protein were found in lysates of E. coli. Besides, the presence of the IPNS gene inhibits expression of a distal gene (galK) in a polycistronic RNA, suggesting that the gene has a low transcriptional efficiency in E. coli.
Patiño
C
C
Departamento de Genética Molecular, Antibióticos S.A., Madrid, Spain.
Sánchez
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F
Peñalva
M A
MA
D000164:Acremonium / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000235:genetics; D004798:Enzymes / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D005786:Gene Expression Regulation; D005800:Genes, Fungal; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D010088:Oxidoreductases; D010957:Plasmids; D011401:Promoter Regions, Genetic
D004271:DNA, Fungal; D004798:Enzymes; D010088:Oxidoreductases; C038370:isopenicillin N synthetase