Nazerian K, Lindahl T, Klein G, Lee L F
J Virol. 1973 Oct;12(4):841-6. doi: 10.1128/JVI.12.4.841-846.1973.
DNA was extracted from [(3)H]thymidine-labeled Marek's disease virus (MDV) and purified by two cycles of CsCl gradient centrifugation in a fixed-angle rotor. The DNA was transcribed in vitro into (32)P-labeled complementary RNA (cRNA). MDV cRNA did not hybridize with DNA from chicken embryo fibroblast cultures or from chicken spleen, but hybridized efficiently with DNA from MDV particles or MDV-infected cell cultures. Five Marek's disease tumors from different chickens and different organs (ovary, liver, testis) were all found to contain MDV DNA sequences. The relative amount of MDV DNA varied from tumor to tumor and was between 3 and 15 virus genome equivalents per cell. The content of virus DNA per cell in spleens from tumor-bearing chickens was much lower than in tumors from the same animals. MDV-infected cell cultures contained a large proportion (28-59%) of virus antigen-positive cells, as measured by immunofluorescence, but tumor cells were negative in this respect (<0.02% positive cells). These data indicate that MDV is present in a provirus form in tumor cells.
从经[³H]胸苷标记的马立克氏病病毒(MDV)中提取DNA,并在固定角度转头中通过两轮氯化铯梯度离心进行纯化。将该DNA体外转录为³²P标记的互补RNA(cRNA)。MDV cRNA不能与鸡胚成纤维细胞培养物或鸡脾脏的DNA杂交,但能与MDV颗粒或MDV感染的细胞培养物的DNA高效杂交。来自不同鸡和不同器官(卵巢、肝脏、睾丸)的5个马立克氏病肿瘤均被发现含有MDV DNA序列。MDV DNA的相对含量因肿瘤而异,每个细胞中为3至15个病毒基因组当量。荷瘤鸡脾脏中每个细胞的病毒DNA含量远低于同一动物肿瘤中的含量。通过免疫荧光检测,MDV感染的细胞培养物中含有很大比例(28 - 59%)的病毒抗原阳性细胞,但肿瘤细胞在这方面呈阴性(阳性细胞<0.02%)。这些数据表明MDV以原病毒形式存在于肿瘤细胞中。