Bacteriophage T4 deoxynucleotide kinase: gene cloning and enzyme purification.
Gene 1 of bacteriophage T4 has been cloned into a lambda pL expression vector, resulting in the overproduction of deoxynucleotide kinase. A procedure that includes affinity chromatography on Cibacron Blue F3GA-agarose has been used to purify milligram quantities of enzymes from a 2-liter culture. The enzyme has been partially characterized in vitro and in vivo, and it appears to be identical to the deoxynucleotide kinase isolated from T4-infected Escherichia coli. These results prove the earlier contention that the phosphorylation of three dissimilar deoxynucleotides (5-hydroxymethyldeoxycytidylate, dTMP, and dGMP), to the exclusion of most others, is catalyzed by a single protein.
Brush
G S
GS
McCollum-Pratt Institute, Johns Hopkins University, Baltimore, Maryland 21218.
Bhatnagar
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SK
Bessman
M J
MJ
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D005816:Genetic Complementation Test; D009703:Nucleoside-Phosphate Kinase / Q000235:genetics* / Q000302:isolation & purification; D010770:Phosphotransferases / Q000235:genetics*; D013379:Substrate Specificity; D013604:T-Phages / Q000201:enzymology* / Q000235:genetics
D010770:Phosphotransferases; D009703:Nucleoside-Phosphate Kinase