Biochemical transformation of thymidine kinase (TK)-deficient mouse cells by herpes simplex virus type 1 DNA fragments purified from hybrid plasmids

Kit, Saul; Otsuka, Hidenori; Qavi, Hamida; Trkula, David; Dubbs, D. R.; Hazen, Marion

doi:10.1093/nar/8.22.5233

Cited by 25 publications

(30 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We disagree with the implications from the work of Colbere-Garapin et al (1979) and more recently Kit et al (1980) that the Sinai site at 0.3045, and especially the 1.2 kb SmaI-EeoRI fragment, define the boundaries of the active HSV-1 TK gene. In our hands, when care was taken to ensure complete digestion, each of these enzymes reduced HSV-1 TK transfer at least 50-fold and in HSV-2 the EcoRI enzyme reduced the efficiency several thousand-fold.…”

Section: Dna)contrasting

confidence: 97%

Transfection with the Isolated Herpes Simplex Virus Thymidine Kinase Genes. I. Minimal Size of the Active Fragments from HSV-1 and HSV-2

Reyes

Jeang

Hayward

1982

Journal of General Virology

View full text Add to dashboard Cite

SUMMARYWe have defined the minimal size and physical map locations in the genomes of both herpes simplex virus types 1 and 2 (HSV-1 and HSV-2) for DNA sequences capable of conferring stable biochemical transformation under thymidine kinase (TK) selection. The experiments involved transfection of Ltk-cells with either isolated virus DNA fragments or cloned pBR322 plasmids containing the 3.5 kilobase (kb) BamHI-O fragment from HSV-I(MP) or the 5.6 kb SalI-G fragment from HSV-2(333). Mapping of restriction enzyme sites within these cloned DNAs, followed by assays for colony formation in HAT medium after transfection with cleaved DNA, localized the biologically active TK-transforming sequences to lie between coordinates 0.300 and 0. Comparison of the putative 5'-end of the HSV-2 TK gene defined by transfection assays, with a 250 bp non-transcribed region at the front of the HSV-1 TK gene, suggests that the promoter regions contain a much higher frequency of conserved cleavage sites than do the coding portions of the two genes. Direct nucleotide sequencing of the 5'-flanking sequences for HSV-2 TK confirmed that large portions of the two promoters possess greater than 95 % sequence homology. At least 140 bp, but no more than 200 bp, of this 5'-promoter region are essential for efficient transfer and expression of the viral TK gene. Combining the results from HSV-1 and HSV-2, we conclude that a contiguous sequence of 1480 to 1540 bp is necessary to achieve at least 10 % of the maximum transformation efficiency.

show abstract

Section: Dna)contrasting

confidence: 97%

Transfection with the Isolated Herpes Simplex Virus Thymidine Kinase Genes. I. Minimal Size of the Active Fragments from HSV-1 and HSV-2

Reyes

Jeang

Hayward

1982

Journal of General Virology

View full text Add to dashboard Cite

show abstract

“…Deletion of sequences within this stretch of 80 nucleotides results in decreased transcription of the tk gene in Xenopus oocytes (McKnight & Gavis, 1980) implying that this sequence is involved in gene recognition by RNA polymerase II. This is consistent with the inactivation of the HSV-1 tk gene by EcoRI cleavage (Wigler et al, 1977;Reyes et al, 1979), although Kit et al (1980) have reported successful transfection with sequences lacking this site.…”

Section: Discussionsupporting

confidence: 83%

Correlation of the Virus Sequence Content and Biological Properties of Cells Carrying the Herpes Simplex Virus Type 2 Thymidine Kinase Gene

Minson¹,

Bell²,

Bastow³

1982

Journal of General Virology

View full text Add to dashboard Cite

SUMMARYSeven cell lines, transformed to a thymidine kinase-positive phenotype with herpes simplex virus type 2 (HSV-2) DNA fragments, have been examined to determine their virus-specific DNA sequence content. The results are consistent with the reported map position of the HSV-2 thymidine kinase gene. Different cell lines contained different amounts of non-selected virus-specific sequences and this correlated with the ability of some cell lines to compensate the deficiencies in some temperature-sensitive (ts) mutants of HSV-1 and HSV-2. The cell lines were also examined for their ability to synthesize elevated levels of thymidine kinase in response to infection with a thymidine kinase-negative virus mutant. Of the seven cell lines, two failed to respond to infection in this way and these cell lines lacked sequences on the upstream side of the kinase gene which were present in the remaining cell lines.

show abstract

“…Transformation of Ltk-cells to the TK+ phenotype with DNA of the cloned HSV-1 TK gene contained in recombinant plasmids or phage was performed under the same conditions as the phage-mediated gene transfer described above. Transformation efficiency comparable to that reported previously (12,38,40) could be attained by this method.…”

supporting

confidence: 89%

“…To establish a method for phage-mediated gene transfer, we used a model system of HSV-1 TK gene and Ltk-cells because this system has been well characterized and is easily available (4,12,25,36,40). We first constructed a recombinant phage carrying cloned HSV-1 TK gene, which also carries the minifibroin gene cloned in pBR322 (Tsujimoto, unpublished data) as a spacer to accommodate the packaging of phage particles (Fig.…”

Section: Resultsmentioning

confidence: 99%

Phage Particle-Mediated Gene Transfer to Cultured Mammalian Cells

Ishiura¹,

Hirose²,

Uchida³

et al. 1982

Mol. Cell. Biol.

View full text Add to dashboard Cite

Recombinant phage particles carrying the thymidine kinase (TK) gene of herpes simplex virus type 1, coprecipitated with calcium phosphate, efficiently transformed mouse Ltk-cells to the TK+ phenotype. The conditions necessary to achieve high efficiency of transfer of the TK gene by phage particle-mediated gene transfer were investigated. Of the parameters examined, the pH of the buffer used for coprecipitation of phage particles with calcium phosphate, the length of time of coprecipitation, and the length of the adsorption period were found to alter the transfer efficiency significantly. The optimal pH was 6.87 at 25°C. Metaphase chromosomes have also been used for transfer of single-copy genes in eucaryotes (2,17,24,26,42) and have at least two advantages over genomic DNA as donors: the efficiency of gene transfer is always 10 to 100 times more than that of genomic DNA, and the transformants are more stable than those obtained by genomic DNA transfer (17). These advantages, we think, are partially due to the organization of the chromosome structure, where DNA sequences are covered with nuclear proteins, packaged compactly into the chromatin structure, and protected from attack by DNase

show abstract

Biochemical transformation of thymidine kinase (TK)-deficient mouse cells by herpes simplex virus type 1 DNA fragments purified from hybrid plasmids

Cited by 25 publications

References 32 publications

Transfection with the Isolated Herpes Simplex Virus Thymidine Kinase Genes. I. Minimal Size of the Active Fragments from HSV-1 and HSV-2

Transfection with the Isolated Herpes Simplex Virus Thymidine Kinase Genes. I. Minimal Size of the Active Fragments from HSV-1 and HSV-2

Correlation of the Virus Sequence Content and Biological Properties of Cells Carrying the Herpes Simplex Virus Type 2 Thymidine Kinase Gene

Phage Particle-Mediated Gene Transfer to Cultured Mammalian Cells

Contact Info

Product

Resources

About