Expression cloning of a sheep adreno-ferredoxin using the polymerase chain reaction.
In addition to the selective amplification of cDNA from total RNA by the PCR method, the distinctive properties of ferredoxin-expressing colonies can be used for cloning a ferredoxin cDNA. This strategy for cloning and expressing cDNA in E. coli was applied to a sheep adreno-ferredoxin. The expressed sheep ferredoxin showed a spectral pattern typical of [2Fe-2S] proteins. The amino acid sequence deduced from the DNA sequence showed that the mature form of sheep ferredoxin consists of 128 amino acid residues. This rapid and simple method for cloning and expressing cDNA can be applied to other ferredoxins.
Matsuo
Y
Y
Department of Biochemistry, Kagawa Medical School, Japan.
Tsujita
M
M
Mizoguchi
K
K
Ichikawa
Y
Y
D000302:Adrenal Cortex; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods*; D004247:DNA; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D005288:Ferredoxins / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D012756:Sheep
D005288:Ferredoxins; D004247:DNA