Identification, cloning, and expression of bacteriophage T5 dnk gene encoding a broad specificity deoxyribonucleoside monophosphate kinase (EC 2.7.4.13).
The nucleotide sequence corresponding to 13-19.5% of the bacteriophage T5 genome in early region C was determined (GenBank AY 140897). One of the five major single-stranded interruptions (nicks) of bacteriophage T5 DNA was identified at 18.5%. The sequenced region was annotated and the putative functions of some open reading frames were proposed by comparison with databases. The dnk gene, encoding a deoxyribonucleoside monophosphate kinase, was identified using a previously defined N-terminal amino acid sequence. The gene was cloned and expressed in Escherichia coli, the enzyme was purified to homogeneity with high yield using two alternative methods, and the recombinant deoxyribonucleoside monophosphate kinase was found to have the same activity and specificity as the native enzyme.
Mikoulinskaia
Galina V
GV
Pushchino Branch of the Institute of Bioorganic Chemistry, Russian Academy of Sciences, Ave. Nauki 6, Pushchino, Moscow region, 142290 Russia.
Zimin
Andrei A
AA
Feofanov
Sergei A
SA
Miroshnikov
Anatolii I
AI
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D015870:Gene Expression; D016679:Genome, Viral; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017856:Phosphotransferases (Phosphate Group Acceptor) / Q000235:genetics* / Q000378:metabolism; D020161:Physical Chromosome Mapping; D010957:Plasmids; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA / Q000379:methods; D013604:T-Phages / Q000235:genetics*
D011994:Recombinant Proteins; D017856:Phosphotransferases (Phosphate Group Acceptor); C020015:(deoxy)nucleoside-phosphate kinase