Quantitative Determination and Location of Newly Synthesized Virus-Specific Ribonucleic Acid in Chicken Cells Infected with Rous Sarcoma Virus

J. Thomas Parsons; John M. Coffin; Richard K. Haroz; Peter A. Bromley; Charles Weissmann

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Quantitative Determination and Location of Newly Synthesized Virus-Specific Ribonucleic Acid in Chicken Cells Infected with Rous Sarcoma Virus

机译：用肉瘤病毒感染鸡细胞新合成病毒特异性核糖核酸的定量测定和位置

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A sensitive and quantitative nucleic acid hybridization assay for the detection of radioactively labeled avian tumor virus-specific RNA in infected chicken cells has been developed. In our experiments we made use of the fact that DNA synthesized by virions of avian myeloblastosis virus in the presence of actinomycin D (AMV DNA) is complementary to at least 35% of the sequences of 70S RNA from the Schmidt-Ruppin strain (SRV) of Rous sarcoma virus. Annealing of radioactive RNA (either SRV RNA or RNA extensively purified from SRV-infected chicken cells) with AMV DNA followed by ribonuclease digestion and Sephadex chromatography yielded products which were characterized as avian tumor virus-specific RNA-DNA hybrids by hybridization competition with unlabeled 70S AMV RNA, equilibrium density-gradient centrifugation in Cs₂SO₄ gradients, and by analysis of their ribonucleotide composition. The amount of viral RNA synthesized during pulse labeling with 3H-uridine could be quantitated by the addition of an internal standard consisting of 32P-labeled SRV RNA prior to purification and hybridization. This quantitative assay was used to determine that, in SRV-infected chicken cells labeled for increasing lengths of time with 3H-uridine, labeled viral RNA appeared first in a nuclear fraction, then in a cytoplasmic fraction, and still later in mature virions. This observation is consistent with the hypothesis that RNA tumor virus RNA is synthesized in the nucleus of infected cells.

机译：已经开发出用于检测感染鸡细胞中放射性标记的禽肿瘤病毒特异性RNA的敏感和定量核酸杂交测定。在我们的实验中，我们利用了在放线霉素D（AMV DNA）存在下的禽类肌卵母细胞素病毒的DNA互补于70 S RNA的至少35％肉瘤病毒的施密特 - 罗普宁菌株（SRV）。用AMV DNA的放射性RNA（广泛纯化的SRV RNA或RNA广泛纯化）和核糖核酸酶消化和Sephadex色谱产生的产物，其特征在于通过杂交竞争与未标记的70次杂交竞争竞争竞争的产物的产物，其特征在于禽肿瘤病毒特异性RNA-DNA杂交物 S AMV RNA，CS _{2 SO _{4 梯度的平衡密度梯度离心，并通过分析其核糖核苷酸组合物。在脉冲标记期间合成的病毒RNA与 3 H-尿苷的量可以通过添加由 32℃的基标标记的SrV RNA组成的内标进行定量，并在纯化之前定量杂交。该定量测定用于确定，在标记的SRV感染的鸡细胞中，标记为增加与 3 H-尿苷的时间长度，首先在核级分中首先出现标记的病毒RNA，然后在细胞质馏分中出现，仍然在成熟的病毒中。该观察结果与在感染细胞核中合成RNA肿瘤病毒RNA的假设一致。}} 展开▼

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《Journal of Virology》 |1973年第5期|共14页

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J. Thomas Parsons; John M. Coffin; Richard K. Haroz; Peter A. Bromley; Charles Weissmann;
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收录信息美国《科学引文索引》(SCI);

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1. Quantitative Determination and Location of Newly Synthesized Virus-Specific Ribonucleic Acid in Chicken Cells Infected with Rous Sarcoma Virus [J] . J. Thomas Parsons, John M. Coffin, Richard K. Haroz, Journal of Virology . 1973,第5期

机译：用肉瘤病毒感染鸡细胞新合成病毒特异性核糖核酸的定量测定和位置

2. Quantitative Determination and Location of Newly Synthesized Virus-Specific Ribonucleic Acid in Chicken Cells Infected with Rous Sarcoma Virus [J] . J. Thomas Parsons, John M. Coffin, Richard K. Haroz, Journal of Virology . 1973,第5期

机译：用肉瘤病毒感染鸡细胞新合成病毒特异性核糖核酸的定量测定和位置

3. Comparison of Rous Sarcoma Virus-Specific Deoxyribonucleic Acid Polymerases in Virions of Rous Sarcoma Virus and in Rous Sarcoma Virus-Infected Chicken Cells [J] . John M. Coffin, Howard M. Temin Journal of Virology . 1971,第5期

机译：Rous Sarcoma病毒特异性脱氧核糖核核酸聚合酶的比较肉瘤病毒和肉瘤病毒感染鸡细胞中的病毒酶

4. Micro-Raman spectroscopy study of ALVAC virus infected chicken embryo cells [C] . Anupam K. Misra, Lori E. Kamemoto, Ningjie Hu, Smart biomedical and physiological sensor technology VIII . 2011

机译：ALVAC病毒感染的鸡胚细胞的显微拉曼光谱研究

5. Identification of Biomarkers Associated with Rous Sarcoma Virus-induced Tumors in Two Divergently Selected Chicken Lines. [D] . Hayden, Ashley. 2016

机译：在两个不同选择的鸡品系中与劳斯肉瘤病毒诱导的肿瘤相关的生物标记物的鉴定。

6. Quantitative Determination and Location of Newly Synthesized Virus-Specific Ribonucleic Acid in Chicken Cells Infected with Rous Sarcoma Virus [O] . J. Thomas Parsons, John M. Coffin, Richard K. Haroz, 1973

机译：新合成病毒特异核糖核酸在劳斯肉瘤病毒感染鸡细胞中的定量测定和定位

7. Cell Cycle-Dependent Activation of Rous Sarcoma Virus-Infected Stationary Chicken Cells: Avian Leukosis Virus Group-Specific Antigens and Ribonucleic Acid [O] . Eric H. Humphries, Howard M. Temin 1972

机译：依赖于肉瘤病毒感染的固定鸡细胞的细胞周期依赖性激活：禽类菌病毒群特异性抗原和核糖核酸

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Quantitative Determination and Location of Newly Synthesized Virus-Specific Ribonucleic Acid in Chicken Cells Infected with Rous Sarcoma Virus

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