Yamamoto K, Mizuno F, Matsuo T, Tanaka A, Nonoyama M, Osato T
Proc Natl Acad Sci U S A. 1978 Oct;75(10):5155-9. doi: 10.1073/pnas.75.10.5155.
Fourteen hybrid clones derived from the fused cultures of human lymphoblastoid FV5 cells and 5-bromodeoxyuridine-resistant mouse fibroblastic MCB2 cells grown in hypoxanthine/aminopterin/thymidine selective medium were examined for the presence of Epstein-Barr virus (EBV) DNA, the expression of the virus-determined nuclear antigen (EBNA), and the presence of human chromosomes, in the course of serial passage in vitro. Among the hybrid clones tested, 3 were positive for EBV DNA and EBNA, whereas the remaining 11 were totally negative. The chromosome investigations showed that human chromosome 14 was consistently involved in all three EBV genome-positive and EBNA-positive hybrid clones, but not in any negative clones. In 10 subclones isolated from 1 of the 3 positive clones, all of which contained only chromosome 14 of the human chromosomes, a concordant segregation of EBNA, EBA DNA, and chromosome 14 was evident. These findings suggest that the resident EBV genome is closely associated with chromosome 14 and the presence of this particular chromosome alone is sufficient for the maintenance and the expression of EBV genetic information in human lymphoblastoid cells.
对在次黄嘌呤/氨基蝶呤/胸腺嘧啶核苷选择性培养基中生长的人淋巴母细胞样FV5细胞和5-溴脱氧尿苷抗性小鼠成纤维细胞MCB2的融合培养物所衍生的14个杂交克隆,在体外连续传代过程中检测了爱泼斯坦-巴尔病毒(EBV)DNA的存在、病毒决定的核抗原(EBNA)的表达以及人类染色体的存在情况。在所测试的杂交克隆中,3个对EBV DNA和EBNA呈阳性,而其余11个完全为阴性。染色体研究表明,人类14号染色体始终存在于所有3个EBV基因组阳性和EBNA阳性的杂交克隆中,但在任何阴性克隆中均未出现。从3个阳性克隆中的1个分离出的10个亚克隆中,所有亚克隆仅含人类染色体中的14号染色体,EBNA、EBA DNA和14号染色体的一致分离很明显。这些发现提示,常驻的EBV基因组与14号染色体密切相关,仅这一特定染色体的存在就足以维持EBV遗传信息在人淋巴母细胞中的表达。