Cloning of coliphage-T4 gene pseT and high-level synthesis of polynucleotide kinase in Escherichia coli.
The gene, pseT, of coliphage T4 which encodes polynucleotide kinase (PNK) was cloned directly into an expression plasmid using the polymerase chain reaction. When placed under the control of the trp promoter, the pse T gene can be maintained stably in Escherichia coli and yields high levels of the enzyme upon induction. The system described facilitates purification and provides very high yields of PNK.
Campos
M
M
Higher Organisms Genetics Division, C.I.G.B., Havana Cuba.
Ortega
M
M
Padrón
G
G
Estrada
M P
MP
de la Fuente
J
J
Herrera
L
L
D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D004274:DNA, Recombinant; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D008969:Molecular Sequence Data; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011116:Polynucleotide 5'-Hydroxyl-Kinase / Q000096:biosynthesis / Q000235:genetics*; D015183:Restriction Mapping; D013604:T-Phages / Q000235:genetics*; D014170:Transformation, Genetic
D004274:DNA, Recombinant; D011116:Polynucleotide 5'-Hydroxyl-Kinase