鼠标细胞的生物化学转化由简单疹病毒DNA碎片DNA
Cell
|May 1, 1977
概括
缺少提米丁激酶 (TK) 的小鼠细胞被设计成通过引入HSV2 DNA片段来表达功能性病毒TK. 这项研究成功地将TK基因映射到HSV基因组上.
科学领域:
- 分子生物学分子生物学
- 病毒学 病毒学
- 遗传学 是一个遗传学.
背景情况:
- 鼠标L细胞缺乏提米丁激酶 (TK),为基因操纵提供了一个模型.
- 简单疹病毒2型 (HSV2) 编码了一个提米丁激酶 (TK) 酶.
研究的目的:
- 确定碎片化HSV2DNA是否可以将TK阳性 (TK+) 现型赋予TK缺乏的小鼠细胞.
- 在病毒基因组上绘制HSV2 TK基因的位置.
主要方法:
- 感染小鼠LMTK细胞与碎片化HSV2DNA (Eco RI,Hind III,或机械剪切).
- 使用HAT介质选择TK+细胞.
- 通过热可溶性和聚烯胺凝电泳分析TK活性.
- 免疫光测试检测HSV特异性抗原.
主要成果:
- 转化细胞获得了TK+表型,表现出病毒特异性的TK活性.
- 转化细胞中的TK活性与感染HSV2.2的细胞中的TK活性相同.
- 转化细胞显示HSV特异性免疫光.
结论:
- 碎片化HSV2DNA可以将小鼠细胞转化为TK+表型.
- 基于这些转换研究,可以绘制HSV2 TK基因在病毒基因组上的位置.
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