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建立含有多个无意义突变和功能抑制tRNA基因的哺乳动物细胞系

R M Hudziak, F A Laski, U L RajBhandary

|November 1, 1982

概括

研究人员创造了具有功能珀抑制基因的稳定哺乳动物细胞系. 这一突破使得特定基因产品的表达成为可能,进步了基因工程和合成生物学应用.

科学领域:

分子生物学分子生物学
遗传学是一个遗传学.
细胞生物学细胞生物学

背景情况:

珀基因突变是过早停止蛋白质合成的代码子.
开发克服珀突变的方法对于基因工程和理解基因功能至关重要.
哺乳动物细胞系是研究基因表达和开发生物技术的重要模型.

研究的目的:

为了产生稳定的哺乳动物细胞系,可以表达来自珀突变基因的蛋白质.
建立一个系统来选择和表征哺乳动物细胞中的珀抑制基因.
为了使哺乳动物系统中工程基因的功能表达.

主要方法:

在简单疹病毒蒂米丁基因酶 (HSV tk),大肠杆菌黄素酸转移酶 (Eco-gpt) 和氨基甘油酸3'转移酶 (NPT-II) 基因中隔离和表征无意义突变.
工程基因具有控制信号,用于大肠杆菌和哺乳动物细胞中的表达.
在哺乳动物细胞中微注射特征无意义突变,以选择珀抑制基因.
在Xenopus laevis tRNA基因的特定位点突变产生珀色抑制基因,然后微注入哺乳动物细胞系.

主要成果:

成功生成了携带功能珀抑制基因的哺乳动物细胞系.
工程细胞系表现出珀突变基因产物的稳定表达.
由此产生的细胞系表现出正常的生长率,表明抑制基因没有不良影响.

结论:

发达的哺乳动物细胞系为表达珀色突变基因提供了一个强大的平台.
这个系统有助于研究基因功能和开发新生物技术工具.
珀抑制基因的稳定整合和表达为合成生物学和基因操纵开辟了新的途径.

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