Sarcoma and transformation-defective viruses produced with infectious DNA(s) from Rous sarcoma virus (RSV)-transformed chicken cells

Hillova, Jana; Dantchev, Dimitre; Mariage, Regine; Plichon, Marie-Pierre; Hill, M.

doi:10.1016/0042-6822(74)90315-8

Cited by 19 publications

(12 citation statements)

References 22 publications

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“…Below this dose the calcium technique using carrier DNA seems to afford the same efficiency of trans fection as the DEAE-dextran does. It is worthy of note that in these trans fections the amount of DNA (0.05 ¡xg/culture) at the end-point dilution is the same as that found in previous studies [8], On the other hand, omission of the carrier DNA in the calcium technique seems to diminish the actual efficiency at the end-point dilution by a factor of 10 as shown in the last column of table I. This means that either with or without carrier DNA the calcium failed to improve the efficiency of the transfection assay over that obtained with the DEAE-dextran, though further assays would be needed to show statistically any minor difference which may exist between these two techniques.…”

supporting

confidence: 67%

“…Care was taken to obtain an equal distribution of DNA solution over the whole surface of the cell monolayer. At the end of this incubation period, 20 ml of Eagle's medium supplemented with 10% tryptose phosphate broth (TPB) and 2% heated calf serum were added, and the cells were grown at 37° under the conditions described [8].…”

mentioning

confidence: 99%

“…51 of chicken cells derived from a fibroblast infected and transformed, in all probability, by one particle of Schmidt-Ruppin RSV, subgroup D (SR-RSV-D). The cloning conditions were described elsewhere [8], The DNA was extracted as described [9] with slight modifications. Briefly, after being washed twice with 0.15 m NaCI -0.1 m EDTA, pH 8.0. the cells were lysed with 0.5% sodium dodecyl sulfate in a buffer containing 0.1 M NaCI, 0.05 M Tris-HCl, pH 8.0, 0.01 M EDTA, and 500 gg/ml pronase.…”

mentioning

confidence: 99%

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Infectivity of Rous Sarcoma Cell DNA: Comparison of Two Techniques of Transfection Assay

Hillova¹,

Hill²,

Goubin³

et al. 1975

Intervirology

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A DNA extracted from a clone of chicken cells transformed by the Schmidt-Ruppin strain of Rous sarcoma virus, subgroup D (SR-RSV-D), was assayed for infectivity by means of DEAE-dextran and calcium techniques. The calcium technique like the previously described DEAE-dextran procedure gave rise to viruses in transfection assays with both native and denatured (SI nuclease susceptible) DNAs. The efficiency of these transfection techniques with native DNA was compared and found to be about the same provided that with the calcium technique carrier DNA was used to complement DNA concentrations lower than 2.5 µg/ml.

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supporting

confidence: 67%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Infectivity of Rous Sarcoma Cell DNA: Comparison of Two Techniques of Transfection Assay

Hillova¹,

Hill²,

Goubin³

et al. 1975

Intervirology

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“…300 were isolated following trans fection as described elsewhere [16). SR-E virus was a recombinant between SR-D No.…”

Section: Cells and Virusesmentioning

confidence: 99%

Tandem Integration of Avian Retroviruses in Double-Infected Cells

Carloni¹,

N²,

Hillova³

et al. 1980

Intervirology

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Chicken embryo fibroblasts were chronically infected by both RAV-1 and SR-E, and established SR-E-transformed quail cells were superinfected with RAV-1. About 75 and 50% of double-infected and superinfected cells, respectively, produced both parental viruses. The DNA purified from these cells contained infectious provirus of both parents and no detectable recombinant provirus. In assays using agar overlay, about 5% of the transformed foci that were induced by the DNA of double-infected, but not superinfected, cells were found to contain the progeny of both parents. This observation suggests that in double infections with avian retroviruses, DNA of both parent viruses may integrate into the host genome either in tandem or in close proximity.

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“…304 and its transformation-defective (td) derivative no. 300 were isolated after transfection of a focuscloned SR-D as described (14). These viruses were grown in Brown Leghorn chf + cells and crude stocks contained, therefore, envE recombinants belonging to subgroup E avian tumor viruses, as indicated by the superscript E. To remove these recombinants SR-D was cloned by triansfection in C/E chfchicken cells, using the DNA purified from SR-D-transformed chf + cells.…”

mentioning

confidence: 99%

Recombinants between avian sarcoma virus genome and chicken helper factor gene of the host cell: cloning by transfection.

Carloni¹,

Kaczorek²,

Hill³

1980

Proc. Natl. Acad. Sci. U.S.A.

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(12) supplemented with 10% (vol/ vol) tryptose phosphate broth (Difco) and 5% heated calf serum. In secondary and later passages of C/E cells, 5% calf serum was sometimes replaced by 5% fetal calf serum. Medium for virus-transformed cultures was supplemented with 1% dimethyl sulfoxide.The presence of the chf gene in chicken cell lines used in this study was assessed by the ability to complement noninfectious Bryan sarcoma virus [BH-RSV(-), in which the helper virus that provides envelope glycoprotein is given in parentheses] released by quail 16Q cells (13). In this assay 10616Q cells were cocultivated with 106 test cells, and 4 days later 5 ml of culture medium was filtered and assayed for infectious pseudotypes in T/B,D turkey cells. In positive assays 20-200 foci of transformed cells formed within 9-12 days, whereas no foci formed under the same conditions in negative assays.Viruses. SR-D no. 304 and its transformation-defective (td) derivative no. 300 were isolated after transfection of a focuscloned SR-D as described (14). These viruses were grown in Brown Leghorn chf + cells and crude stocks contained, therefore, envE recombinants belonging to subgroup E avian tumor viruses, as indicated by the superscript E. To remove these recombinants SR-D was cloned by triansfection in C/E chfchicken cells, using the DNA purified from SR-D-transformed chf + cells. In this proviral-DNA-cloning procedure the DNA was serially diluted and the virus was isolated from a culture transfected at the end-point dilution of the DNA, thus with all probability generated from the provirus(es) expressed in one transfected cell.Cloned SR-D was multiplied in C/E chf-cells grown in Corning 490-cm2 plastic roller bottles. Culture medium was harvested twice a day for several days, centrifuged at 10,000 X g and 4°C for 15 min, and stored at -700 C.

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Sarcoma and transformation-defective viruses produced with infectious DNA(s) from Rous sarcoma virus (RSV)-transformed chicken cells

Cited by 19 publications

References 22 publications

Infectivity of Rous Sarcoma Cell DNA: Comparison of Two Techniques of Transfection Assay

Infectivity of Rous Sarcoma Cell DNA: Comparison of Two Techniques of Transfection Assay

Tandem Integration of Avian Retroviruses in Double-Infected Cells

Recombinants between avian sarcoma virus genome and chicken helper factor gene of the host cell: cloning by transfection.

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