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转化细胞中感染性DNA的证明。III. 感染性DNA-蛋白质复合物的检测与猿猴腺病毒SA7诱导的肿瘤细胞中病毒持续性的相关性。

Demonstration of infectious DNA in transformed cells. III. Correlation of detection of infectious DNA-protein complexes with persistence of virus in simian adenovirus SA7-induced tumor cells.

作者信息

Butel J S, TAlas M, Ugur J, Melnick J L

出版信息

Intervirology. 1975;5(1-2):43-56. doi: 10.1159/000149879.

Abstract

Primary tumors induced in newborn hamsters by simian adenovirus SA7 were investigated in transfection experiments. Infectious DNA-protein complexes were readily detected in both the supernatant and pellet fractions obtained by a modified Hirt extraction procedure; DEAE-dextran was required for infectivity to become manifest. Infectivity could be abolished by exposure to DNase, but it was unaffected by SA7-specific antiserum or RNase, and only partially inactivated by trypsin treatment. SA7 tumor cells serially passaged either in tissue culture or in hamsters yielded infectious DNA complexes much less frequently and appeared to evolve into nonyielder cell lines. When large numbers of cells were lysed, intact virus could be recovered from all the primary tumors and from some of the subcultured cell lines. There was a correlation between the persistence of complete virus and the presence of infectious DNA-protein complexes. When the tumor cells carried very small amounts of intact virus, infectious DNA complexes could still be detected; when virus could no longer be detected in the tumor cells, infectious DNA complexes could no longer be found. The results suggest that a portion of the infectious DNA moieties exists as viral DNA-protein complexes in the tumor cells.

摘要

在转染实验中对新生仓鼠中由猿猴腺病毒SA7诱导产生的原发性肿瘤进行了研究。通过改良的Hirt提取程序获得的上清液和沉淀部分中均易于检测到感染性DNA-蛋白质复合物;感染性的显现需要DEAE-葡聚糖。感染性可通过暴露于DNase而消除,但不受SA7特异性抗血清或RNase的影响,且仅经胰蛋白酶处理会部分失活。在组织培养或仓鼠体内连续传代的SA7肿瘤细胞产生感染性DNA复合物的频率要低得多,并且似乎演变成了不产生感染性的细胞系。当大量细胞裂解时,可从所有原发性肿瘤和一些传代培养的细胞系中回收完整病毒。完整病毒的持续存在与感染性DNA-蛋白质复合物的存在之间存在相关性。当肿瘤细胞携带极少量完整病毒时,仍可检测到感染性DNA复合物;当在肿瘤细胞中不再能检测到病毒时,也不再能找到感染性DNA复合物。结果表明,一部分感染性DNA部分以病毒DNA-蛋白质复合物的形式存在于肿瘤细胞中。

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