Pater A, Pater M M
Basic Medical Sciences, Faculty of Medicine, Memorial University of Newfoundland, St. John's, Canada.
Virology. 1988 Apr;163(2):625-8. doi: 10.1016/0042-6822(88)90305-4.
Transformation of primary human embryonic kidney (HEK) cells as selected by either focus assay or growth in soft agar after their transfection with BK virus (BKV) DNA alone or BKV DNA plus the activated form of human Ha-ras oncogene has been previously reported (A. Pater and M. M. Pater, J. Virol. 58, 680-683 (1986]. In order to examine the expression of the regulatory elements of papovaviruses in each of the transformed phenotypes, chloramphenical acetyltransferase (CAT) plasmids under the control of BK or SV40 early promotor were used in transient assays. The expression of CAT, as driven by either SV40 or BK early promoters, was approximately 100-fold higher in HEK cells transformed by BKV DNA plus Ha-ras oncogene than in cells transformed only by BKV DNA. The higher CAT expression was not due to high levels of plasmid replication in these cells. Time course studies revealed that the higher CAT activity could be explained largely by greater plasmid uptake and stability in BK plus ras-transformed cells.
先前已有报道，单独用BK病毒（BKV）DNA或BKV DNA加上人Ha-ras癌基因的激活形式转染原代人胚肾（HEK）细胞后，通过病灶分析或软琼脂培养筛选出的细胞发生了转化（A. 佩特和M. M. 佩特，《病毒学杂志》58，680 - 683（1986年））。为了检测乳头瘤病毒调节元件在每种转化表型中的表达，在瞬时分析中使用了受BK或SV40早期启动子控制的氯霉素乙酰转移酶（CAT）质粒。由SV40或BK早期启动子驱动的CAT在经BKV DNA加Ha-ras癌基因转化的HEK细胞中的表达比仅经BKV DNA转化的细胞高约100倍。较高的CAT表达并非由于这些细胞中质粒的高复制水平。时间进程研究表明，较高的CAT活性很大程度上可以通过BK加ras转化细胞中更高的质粒摄取和稳定性来解释。
