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转化DNA的分离:克隆仓鼠子基因

I Lowy, A Pellicer, J F Jackson

|December 1, 1980

概括

研究人员通过基因转移和分子克隆分离了仓鼠腺酸转移酶 (aprt) 基因. 这种方法成功地识别和分离了aprt基因,用于隔离其他功能基因的潜在应用.

科学领域:

分子生物学分子生物学
遗传学是一个遗传学.
生物化学生物化学

背景情况:

腺因基转移酶 (aprt) 酶在纯素代谢中起着至关重要的作用.
隔离特定的基因对于理解基因功能和开发生物技术应用至关重要.

研究的目的:

为了分离编码腺酸转移酶 (aprt) 的仓鼠基因.
开发一种方法,利用基因转移和分子克隆来分离功能性基因.

主要方法:

将与pBR322DNA相结合的Hind III分裂仓鼠基因组DNA转移到小鼠细胞中.
使用羊羔重组菌体库进行aprt基因的分子克隆.
对转化细胞DNA进行同源序列分析.

主要成果:

成功地将小鼠aprt-细胞转化为aprt+表型,表明成功的基因转移.
隔离了一种含有与pBR322DNA相关的仓鼠 aprt基因的羔羊重组菌体.
证实了所有仓鼠 aprt+变形体中与分离的克隆相同的序列的存在.

结论:

这项研究成功地分离了使用基因转移和分子克隆的组合来分离仓鼠子基因.
开发的实验设计在理论上适用于通过DNA介导的基因转移来分离任何具有可选择或可识别功能的基因.

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In-vitro Mutagenesis

In-vitro Mutagenesis

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.