Gene/Protein
Disease
Symptom
Drug
Enzyme
Compound
Pivot Concepts:
Gene/Protein
Disease
Symptom
Drug
Enzyme
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Target Concepts:
Gene/Protein
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Query: EC:2.7.1.21 (
thymidine kinase
)
7,561
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
Yeast plasmids have been constructed that carry the Herpes simplex Virus type 1 (HSV-1)
thymidine kinase
(TK) gene which is functionally expressed in Saccharomyces cerevisiae. The expression of the TK gene appears to be due to transcriptional read-through from a yeast promoter that lies on the 3' side of the
HIS3
gene. The TK+ yeast possesses in vitro
thymidine kinase
activity which is absent in the original yeast strain. Yeast strains auxotrophic for thymidine monophosphate (dTMP) (tmp1) can grown on thymidine-containing medium after transformation with these plasmids. Tmp+, TK+ S. cerevisiae whose de novo synthesis of dTMP is inhibited with amethopterin plus sufanilamide is also capable of growth in thymidine. S. cerevisiae transformed with such plasmids is capable of incorporating thymidine and bromodeoxyuridine into DNA.
...
PMID:Expression of the Herpes simplex virus thymidine kinase gene in Saccharomyces cerevisiae. 627 52
The thymidine analogue BrdU is a powerful tool for analysing nucleotide incorporation in studies of DNA replication or repair. S. cerevisiae lacks the thymidine salvage pathway that enables efficient cellular uptake and incorporation of thymidine analogues into DNA. Recent in vivo reconstitution of this pathway in yeast by high-level expression of Herpes simplex virus
thymidine kinase
(HSV-TK) or combined expression of HSV-TK and human equilibrative nucleoside transporter (hENT1) has enabled analysis of BrdU incorporation in yeast. While the BrdU incorporation systems are highly valuable, the construction and use of strains utilizing these systems can be complicated by specific requirements of the available systems. We have created a set of vectors that simplify the construction and use of BrdU-Incorporating (BrdU-Inc) strains of budding yeast. Each vector in the set contains HSV-TK and hENT1 under the control of promoters that express constitutively, and one of four different selectable markers (
HIS3
, TRP1, LEU2 or URA3) for genomic integration. With these BrdU-Inc vectors, one-step integration of a single copy produces yeast that efficiently incorporate BrdU upon its addition to the medium. These vectors ease strain construction and maintenance, thereby facilitating routine use of BrdU for analysis in yeast.
...
PMID:New vectors for simplified construction of BrdU-Incorporating strains of Saccharomyces cerevisiae. 1708 35