Schneider-Gädicke A, Kaul S, Schwarz E, Gausepohl H, Frank R, Bastert G
Institut für Virusforschung, Deutsches Krebsforschungszentrum, Heidelberg, Federal Republic of Germany.
Cancer Res. 1988 Jun 1;48(11):2969-74.
We recently reported the transcription patterns of human papillomavirus (HPV) type 18 sequences in human cervical carcinoma cell lines. The open reading frames (ORFs) E6* and E6 represent the 5'-terminal cistrons in HPV18 mRNAs. ORF E6* was assumed to be specific for HPV types associated with genital carcinomas. To identify the predicted gene product, ORF E6* from a HeLa cDNA clone was expressed as an MS2 fusion protein in Escherichia coli. The C-terminal 23 amino acid residues were chemically synthesized. A panel of monoclonal antibodies was generated, recognizing E6* and E6* plus E6, respectively. In human cervical carcinoma cell lines grown in vitro these monoclonal antibodies specifically immunoprecipitate the putative Mr 17,000 and 18,000 HPV18 E6 proteins in nuclear protein fractions. In a HPV18 DNA containing human cervical carcinoma established in nude mice, these monoclonal antibodies specifically immunoprecipitate a polypeptide with a molecular weight of 6500 as predicted for the HPV18 ORF E6* gene product in a nuclear protein fraction.
我们最近报道了人乳头瘤病毒18型（HPV18）序列在人宫颈癌细胞系中的转录模式。开放阅读框（ORF）E6和E6是HPV18 mRNA中的5'-末端顺反子。ORF E6被认为是与生殖器癌相关的HPV类型所特有的。为了鉴定预测的基因产物，将来自HeLa cDNA克隆的ORF E6在大肠杆菌中表达为MS2融合蛋白。C末端的23个氨基酸残基通过化学合成获得。制备了一组单克隆抗体，分别识别E6以及E6加E6。在体外培养的人宫颈癌细胞系中，这些单克隆抗体能特异性免疫沉淀核蛋白组分中推定的分子量为17,000和18,000的HPV18 E6蛋白。在裸鼠中建立的含HPV18 DNA的人宫颈癌模型中，这些单克隆抗体能特异性免疫沉淀核蛋白组分中分子量为6500的一种多肽，这与HPV18 ORF E6基因产物的预测分子量相符。
