Cloning, sequence analysis and overexpression of a rhodothermus marinus gene encoding a thermostable thymidine kinase.
Thymidine kinase type II is an important part of the pyrimidine salvage pathway. The thymidine kinase gene from the thermophilic eubacterium Rhodothermus marinus was cloned, sequenced and overexpressed. The gene is 639 bp and encodes a protein of 213 amino acids with a calculated molecular mass of 23.6 kDa. It shows homology to other thymidine kinase proteins from eukaryotic and prokaryotic organisms. The recombinant protein is inhibited by dNTPs but not by dNDPs. It is a tetramer in its native state. Its optimum temperature of activity is 65 degrees C and it has a half life of 15 min at 90 degrees C. This is the first thymidine kinase to be described from a thermophilic bacterium.
Blöndal
T
T
Institute of Biology, University of Iceland, IS-108, Reykjavik, Iceland.
Thorbjarnardóttir
S H
SH
Kieleczawa
J
J
Einarsson
J M
JM
Hjörleifsdóttir
S
S
Kristjánsson
J K
JK
Eggertsson
G
G
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D005798:Genes, Bacterial; D006090:Gram-Negative Bacteria / Q000201:enzymology / Q000235:genetics*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011994:Recombinant Proteins / Q000096:biosynthesis; D013937:Thymidine Kinase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
D011994:Recombinant Proteins; D013937:Thymidine Kinase