应用重组质粒在大肠杆菌中表达人乳头瘤病毒(HPV)16 型 E7 基因.以所产生的 E7 融合蛋白为抗原免疫家兔,制得抗 E7 蛋白抗血清.在子宫颈癌组织切片中用此抗血清作免疫组化染色(胶体金标记染色法).在光学显微镜下可观察到癌细胞中存在 E7 抗原黑色颗粒,位于细胞核内.主要附着于核膜,可证明 E7 基因在 HPV16 感染的子宫颈癌细胞中有强烈表达;提示 E7 基因可能即为 HPV16的癌基因.
The E7 gene of human papillomavirus type 16 was expressed in Escherichia coli with a recombinant plasmid prepared previously in our laboratory.The E7 fusion protein produced and purified by gel electrophoresis was used as antigen to immunize rabbits for preparing anti-E7 protein antisera.The antisera were used to stain the tissue sections often cases of uterine cervical cancer with an immunohistochemical technique(colloidal gold labeled stain).The E7 antigen was seen in the cells of 6 cases of cervical carcinoma as black granules under light microscope. The E7 protein was found in the nucleus of positive cells, mainly on the nuclear membrane, arranged in a circle. Only a few silver stained granules were seen in the cytoplasm. We believe that the intracellular distribution of the E7 protein indicates that there is high expression of the E7 gene in the HPV infected cells of the cervical cancer; and the fact that E7 antigen is a nuclear protein suggests that E7 gene is an oncogene of HPV 16.
徐钤,齐凤菊.人乳头瘤病毒16型 E7蛋白在子宫颈癌细胞内的定位[J].生物化学与生物物理进展,1993,20(2):108-111
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