Transfer of herpes simplex virus thymidine kinase synthesized in bacteria by a high-expression plasmid to tissue culture cells by protoplast fusion
The introduction of a protein into living tissue culture cells may permit the in vivo study of functions of the protein. The authors have previously described a high-efficiency-expression plasmid, pHETK2, containing the herpes simplex virus type 1 thymidine kinase (TK) gene which, upon temperature induction, causes TK to be synthesized as greater than 4% of the bacterial protein. In this report it is shown that enzymatically active TK was transferred to mouse Ltk- cells by polyethylene glycol-mediated fusion with protoplasts prepared from bacteria containing induced levels of TK. The presence of TK in the Ltk- cells was detected by the incorporation of (/sup 3/H)thymidine into cell nuclei as measured by autoradiography.
- Research Organization:
- Johns Hopkins Univ., Baltimore, MD
- OSTI ID:
- 5947627
- Journal Information:
- Mol. Cell. Biol.; (United States), Vol. 4:8
- Country of Publication:
- United States
- Language:
- English
Similar Records
Phenotypic switching in cells transformed with the herpes simplex virus thymidine kinase gene
Optimized thymidylate kinase assay, based on enzymatically synthesized 5-(/sup 125/I)iododeoxyuridine monophosphate and its application to an immunological study of herpes simplex virus thymidine-thymidylate kinases
Related Subjects
ENZYMES
BIOLOGICAL FUNCTIONS
THYMIDINE
AUTORADIOGRAPHY
BIOSYNTHESIS
CELL CULTURES
CELL NUCLEI
ESCHERICHIA COLI
GENES
HERPES SIMPLEX
MICE
PLASMIDS
PROTEINS
TRITIUM COMPOUNDS
ANIMALS
AZINES
BACTERIA
CELL CONSTITUENTS
DISEASES
FUNCTIONS
HETEROCYCLIC COMPOUNDS
INFECTIOUS DISEASES
LABELLED COMPOUNDS
MAMMALS
MICROORGANISMS
NUCLEOSIDES
NUCLEOTIDES
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PYRIMIDINES
RIBOSIDES
RODENTS
SKIN DISEASES
SYNTHESIS
VERTEBRATES
VIRAL DISEASES
550201* - Biochemistry- Tracer Techniques