title: Isolation and overexpression in Escherichia coli of the flavodoxin gene from Anabaena PCC 7119.
abstract: The gene coding for flavodoxin from Anabaena PCC 7119 was cloned by using the polymerase chain reaction (PCR). The gene is transcribed into a 1250-base transcript. The expression of the flavodoxin gene was analysed and found to be regulated at the transcriptional level by the availability of iron. The PCR-amplified gene was cloned into the expression vector pTrc 99b and expressed in Escherichia coli. High concentrations of flavodoxin were found (20% of total protein). The recombinant protein was purified from the cytosolic fraction of the cells and it exhibited properties identical with those of the wild-type Anabaena flavodoxin.
authors.affiliation: Department of Molecular Cell Biology, University of Utrecht, The Netherlands.
authors.forename: M F
authors.initials: MF
authors.lastname: Fillat
authors.forename: W E
authors.initials: WE
authors.lastname: Borrias
authors.forename: P J
authors.initials: PJ
authors.lastname: Weisbeek
mesh_terms: D000595:Amino Acid Sequence; D017033:Anabaena / Q000235:genetics*; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D005418:Flavodoxin / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D015870:Gene Expression; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016133:Polymerase Chain Reaction / Q000379:methods; D012329:RNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification
chemical_list: D004269:DNA, Bacterial; D005418:Flavodoxin; D012329:RNA, Bacterial; D011994:Recombinant Proteins