scholarly journals Transfection with the Isolated Herpes Simplex Virus Thymidine Kinase Genes. II. Evidence for Amplification of Viral and Adjacent Cellular DNA Sequences

1982 ◽  
Vol 60 (2) ◽  
pp. 209-224 ◽  
Author(s):  
G. R. Reyes ◽  
M. W. McLane ◽  
G. S. Hayward
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Thymidine Kinase ◽  
Dna Sequences ◽  
Cellular Dna ◽  
Simplex Virus

scholarly journals Adaptation of a retrovirus as a eucaryotic vector transmitting the herpes simplex virus thymidine kinase gene.

1982 ◽  
Vol 2 (4) ◽  
pp. 426-436 ◽  
Author(s):  
C J Tabin ◽  
J W Hoffmann ◽  
S P Goff ◽  
R A Weinberg
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Thymidine Kinase ◽  
Dna Sequences ◽  
Leukemia Virus ◽  
Chimeric Virus ◽  
Animal Cells ◽  
Kinase Gene ◽  
General Applicability ◽  
Simplex Virus

We investigated the feasibility of using retroviruses as vectors for transferring DNA sequences into animal cells. The thymidine kinase (tk) gene of herpes simplex virus was chosen as a convenient model. The internal BamHI fragments of a DNA clone of Moloney leukemia virus (MLV) were replaced with a purified BamHI DNA segment containing the tk gene. Chimeric genomes were created carrying the tk insert in both orientations relative to the MLV sequence. Each was transfected into TK- cells along with MLV helper virus, and TK+ colonies were obtained by selection in the presence of hypoxanthine, aminopterin, and thymidine (HAT). Virus collected from TK+-transformed, MLV producer cells passed the TK+ phenotype to TK- cells. Nonproducer cells were isolated, and TK+ transducing virus was subsequently rescued from them. The chimeric virus showed single-hit kinetics in infections. Virion and cellular RNA and cellular DNA from infected cells were all shown to contain sequences which hybridized to both MLV- and tk-specific probes. The sizes of these sequences were consistent with those predicted for the chimeric virus. In all respects studied, the chimeric MLV-tk virus behaved like known replication-defective retroviruses. These experiments suggest great general applicability of retroviruses as eucaryotic vectors.

scholarly journals Satellite DNA induces unstable expression of the adjacent herpes simplex virus tk gene cotransfected in mouse cells.

1988 ◽  
Vol 8 (3) ◽  
pp. 1336-1344 ◽  
Author(s):  
D Talarico ◽  
A F Peverali ◽  
E Ginelli ◽  
R Meneveri ◽  
C Mondello ◽  
...  
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Dna Sequences ◽  
Satellite Dna ◽  
Repetitive Sequences ◽  
Metaphase Chromosomes ◽  
Highly Repetitive Dna ◽  
Cellular Dna ◽  
Mouse Satellite Dna ◽  
Simplex Virus

To study the influence of clustered highly repetitive DNA sequences on the expression of adjacent genes, LTK- cells were cotransfected with the herpes simplex virus thymidine kinase (tk) gene and mouse satellite DNA. TK+ transformants containing a few copies of the tk genes flanked by satellite DNA were isolated. In situ hybridization on the metaphase chromosomes indicated that in each cell line the TK sequences resided at a single chromosomal site and that integration occurred preferentially into regions of the cellular DNA rich in highly repetitive sequences. The prominent feature of these cell lines was their phenotypic instability. Suppression and reexpression of the tk gene occurred at high frequency (greater than 3%) and did not correlate with any significant change in the organization of foreign DNA or with the presence of selective agents. These results indicate that satellite DNA, the major component of constitutive heterochromatin, may influence the expression of adjacent genes by affecting the chromatin structure.

Satellite DNA induces unstable expression of the adjacent herpes simplex virus tk gene cotransfected in mouse cells

1988 ◽  
Vol 8 (3) ◽  
pp. 1336-1344
Author(s):  
D Talarico ◽  
A F Peverali ◽  
E Ginelli ◽  
R Meneveri ◽  
C Mondello ◽  
...  
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Dna Sequences ◽  
Satellite Dna ◽  
Repetitive Sequences ◽  
Metaphase Chromosomes ◽  
Highly Repetitive Dna ◽  
Cellular Dna ◽  
Mouse Satellite Dna ◽  
Simplex Virus

To study the influence of clustered highly repetitive DNA sequences on the expression of adjacent genes, LTK- cells were cotransfected with the herpes simplex virus thymidine kinase (tk) gene and mouse satellite DNA. TK+ transformants containing a few copies of the tk genes flanked by satellite DNA were isolated. In situ hybridization on the metaphase chromosomes indicated that in each cell line the TK sequences resided at a single chromosomal site and that integration occurred preferentially into regions of the cellular DNA rich in highly repetitive sequences. The prominent feature of these cell lines was their phenotypic instability. Suppression and reexpression of the tk gene occurred at high frequency (greater than 3%) and did not correlate with any significant change in the organization of foreign DNA or with the presence of selective agents. These results indicate that satellite DNA, the major component of constitutive heterochromatin, may influence the expression of adjacent genes by affecting the chromatin structure.

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