Novak U, Dilworth S M, Griffin B E
Proc Natl Acad Sci U S A. 1980 Jun;77(6):3278-82. doi: 10.1073/pnas.77.6.3278.
Rat-1 cells were transfected with the restriction enzyme fragment of polyoma virus DNA that extends clockwise from the Bcl I site ((65.4 map units) to the EcoRI site (0/100 map units). Six transformed cell lines were obtained and one of them (BE-1) has been investigated in detail. The viral DNA that is integrated into host DNA in this line appeared to consist of two fragments arranged in a "head-to-tail" tandem with no detectable intervening host sequences. BE-1 cells contained polyoma virus small and middle tumor antigens that were indistinguishable from the corresponding tumor antigens from lytically infected cells. No large tumor antigen was detected but a "new" Mr 34,000 protein, which proved to be a truncated version of large tumor antigen, was immunoprecipitated by anti-tumor-antigen antiserum. After injection of 10(6) BE-1 cells into young syngeneic Fischer rats, tumors appeared within 3--4 weeks. Thus, the coding capacity of the Bcl I/EcoRI fragment of polyoma virus DNA is sufficient to enable the cells to produce all of small and middle tumor antigens and about a third of large tumor antigen, to transform cells stably in culture, and to produce tumors in vivo.
用多瘤病毒DNA的限制性酶切片段转染Rat-1细胞,该片段从Bcl I位点(65.4个图谱单位)顺时针延伸至EcoRI位点(0/100个图谱单位)。获得了六个转化细胞系,其中一个(BE-1)已被详细研究。整合到该细胞系宿主DNA中的病毒DNA似乎由两个片段组成,呈“头对头”串联排列,未检测到中间的宿主序列。BE-1细胞含有多瘤病毒小肿瘤抗原和中肿瘤抗原,它们与来自裂解感染细胞的相应肿瘤抗原无法区分。未检测到大肿瘤抗原,但一种“新的”分子量为34,000的蛋白质被抗肿瘤抗原抗血清免疫沉淀,该蛋白质被证明是大肿瘤抗原的截短版本。将10⁶个BE-1细胞注射到同基因的年轻Fischer大鼠体内后,3至4周内出现了肿瘤。因此,多瘤病毒DNA的Bcl I/EcoRI片段的编码能力足以使细胞产生所有的小肿瘤抗原和中肿瘤抗原以及约三分之一的大肿瘤抗原,在培养中稳定转化细胞,并在体内产生肿瘤。