Transfer of herpes simplex virus thymidine kinase synthesized in bacteria by a high-expression plasmid to tissue culture cells by protoplast fusion

Journal Article · Wed Aug 01 04:00:00 EDT 1984 · Mol. Cell. Biol.; (United States)

DOI:https://doi.org/10.1128/MCB.4.8.1644· OSTI ID:5947627

Waldman, A S; Milman, G

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), Journal Name: Mol. Cell. Biol.; (United States) Vol. 4:8; ISSN MCEBD
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
AUTORADIOGRAPHY
AZINES
BACTERIA
BIOLOGICAL FUNCTIONS
BIOSYNTHESIS
CELL CONSTITUENTS
CELL CULTURES
CELL NUCLEI
DISEASES
ENZYMES
ESCHERICHIA COLI
FUNCTIONS
GENES
HERPES SIMPLEX
HETEROCYCLIC COMPOUNDS
INFECTIOUS DISEASES
LABELLED COMPOUNDS
MAMMALS
MICE
MICROORGANISMS
NUCLEOSIDES
NUCLEOTIDES
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PLASMIDS
PROTEINS
PYRIMIDINES
RIBOSIDES
RODENTS
SKIN DISEASES
SYNTHESIS
THYMIDINE
TRITIUM COMPOUNDS
VERTEBRATES
VIRAL DISEASES