Krippl B, Andrisani O, Jones N, Westphal H, Rosenberg M, Ferguson B
J Virol. 1986 Aug;59(2):420-7. doi: 10.1128/JVI.59.2.420-427.1986.
We efficiently expressed, in Escherichia coli, and purified the protein product encoded by the human adenovirus type 12 (Ad12) 13S mRNA. The functional properties of the E1A protein were analyzed by introducing the protein by microinjection or protoplast fusion into living mammalian cells. We showed that the E. coli-expressed E1A protein induces gene expression of the adenovirus type 5 (Ad5) E1A deletion mutant Ad5dl312. The purified E1A protein rapidly and quantitatively localized to the cell nucleus after microinjection into the cytoplasm. In addition, we raised high-titered monospecific antibodies to the purified Ad12 E1A protein. Using deleted forms of an adenovirus type 2 and Ad5 hybrid (Ad2/5) E1A protein, we showed that all of the epitopes conserved between Ad2/5 E1A and Ad12 E1A protein that are recognized by the Ad12 E1A-specific antiserum map to within the first exon-encoded amino-terminal half of the protein.
我们在大肠杆菌中高效表达并纯化了人12型腺病毒（Ad12）13S mRNA编码的蛋白质产物。通过显微注射或原生质体融合将该蛋白质导入活的哺乳动物细胞，对E1A蛋白的功能特性进行了分析。我们发现，大肠杆菌表达的E1A蛋白可诱导5型腺病毒（Ad5）E1A缺失突变体Ad5dl312的基因表达。将纯化的E1A蛋白显微注射到细胞质后，它能迅速且定量地定位于细胞核。此外，我们制备了针对纯化的Ad12 E1A蛋白的高滴度单特异性抗体。利用2型腺病毒和Ad5杂交体（Ad2/5）E1A蛋白的缺失形式，我们发现Ad12 E1A特异性抗血清识别的Ad2/5 E1A和Ad12 E1A蛋白之间所有保守表位都位于该蛋白第一个外显子编码的氨基末端一半范围内。
