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表达5型腺病毒前体末端蛋白的293和HeLa细胞系。

Adenovirus type 5 precursor terminal protein-expressing 293 and HeLa cell lines.

作者信息

Schaack J, Guo X, Ho W Y, Karlok M, Chen C, Ornelles D

机构信息

Department of Microbiology, University of Colorado Health Sciences Center, Denver 80262, USA.

出版信息

J Virol. 1995 Jul;69(7):4079-85. doi: 10.1128/JVI.69.7.4079-4085.1995.

Abstract

HeLa and 293 cell lines that express biologically active adenovirus type 5 precursor terminal protein (pTP) have been made. The amount of pTP synthesized in these cell lines ranges from barely detectable to greater than that observed in cells infected with the wild-type virus. The pTP-expressing cell lines permit the growth of a temperature-sensitive terminal protein mutant virus sub100r at the nonpermissive temperature. A higher percentage of the stably transfected 293 cell lines expressed terminal protein, and generally at considerably higher levels, than did the HeLa cell lines. While 293 cells appeared to tolerate pTP better than did HeLa cells, high-level pTP expression in 293 cells led to a significantly reduced growth rate. The 293-pTP cell lines produce infectious virus after transfection with purified viral DNA and form plaques when overlaid with Noble agar after infection at low multiplicity. These cell lines offer promise for the production of adenoviruses lacking pTP expression and therefore completely defective for replication.

摘要

已构建出表达具有生物活性的5型腺病毒前体末端蛋白(pTP)的HeLa和293细胞系。在这些细胞系中合成的pTP量从几乎检测不到到高于感染野生型病毒的细胞中观察到的量不等。表达pTP的细胞系允许温度敏感型末端蛋白突变病毒sub100r在非允许温度下生长。与HeLa细胞系相比,更高比例的稳定转染293细胞系表达末端蛋白,并且通常表达水平相当高。虽然293细胞似乎比HeLa细胞更能耐受pTP,但293细胞中高水平的pTP表达导致生长速率显著降低。293-pTP细胞系在用纯化的病毒DNA转染后产生感染性病毒,并且在低 multiplicity感染后用Noble琼脂覆盖时形成噬斑。这些细胞系有望用于生产缺乏pTP表达因而完全复制缺陷的腺病毒。

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