从酵母人工染色体克隆图书馆中分离出单复制人类基因
B H Brownstein1, G A Silverman, R D Little
1Department of Genetics, Washington University School of Medicine, St. Louis, MO 63110.
概括
研究人员用酵母人造染色体克隆人类基因. 这种方法允许大DNA片段的稳定传播,为克隆整个人类基因组铺平了道路.
科学领域:
- 遗传学 是一个遗传学.
- 分子生物学分子生物学
- 酵母遗传学 酵母遗传学
背景情况:
- 一个新的克隆系统利用酵母人造染色体 (YAC) 来传播大型DNA分子.
- 该系统为克隆大型DNA段,包括人类基因组部分提供了潜在的方法.
- 从YAC库中有效地恢复和传播特定序列对应用程序至关重要.
研究的目的:
- 用YAC系统证明克隆特定人类基因的可行性.
- 在YAC中评估克隆人类DNA序列的稳定性和忠实性.
- 为了分离和表征含有特定单拷贝人类基因的YAC克隆.
主要方法:
- 从全人类DNA构建一个YAC库.
- 选YAC图书馆的特定人类基因.
- 对于大小和稳定性来说,孤立的YAC克隆的特征.
主要成果:
- 两个单复制的人类基因,因子IX和等离子素激活剂抑制剂2型,已成功克隆.
- 对于每个基因,获得了多个独立的YAC分离物.
- 克隆的DNA片段在60至650千基之间,在酵母中长期繁殖时稳定.
结论:
- 酵母人造染色体系统对于克隆大型人类DNA片段是有效的.
- 含有人类基因的YAC克隆是稳定的,并且似乎是忠实的复制品.
- 这项技术对大规模的基因组克隆工作具有前景.
相关概念视频
Recombinant DNA
Overview
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.
Reproductive Cloning
Reproductive cloning is the process of producing a genetically identical copy—a clone—of an entire organism. While clones can be produced by splitting an early embryo—similar to what happens naturally with identical twins—cloning of adult animals is usually done by a process called somatic cell nuclear transfer (SCNT).
Somatic Cell Nuclear Transfer
In SCNT, an egg cell is taken from an animal and its nucleus is removed, creating an enucleated egg. Then a somatic cell—any cell that is not a sex...
Somatic Cell Nuclear Transfer
In SCNT, an egg cell is taken from an animal and its nucleus is removed, creating an enucleated egg. Then a somatic cell—any cell that is not a sex...
Reproductive Cloning
Reproductive cloning is the process of producing a genetically identical copy—a clone—of an entire organism. While clones can be produced by splitting an early embryo—similar to what happens naturally with identical twins—cloning of adult animals is usually done by a process called somatic cell nuclear transfer (SCNT).
Somatic Cell Nuclear Transfer
In SCNT, an egg cell is taken from an animal and its nucleus is removed, creating an enucleated egg. Then a somatic cell—any cell that is not a sex...
Somatic Cell Nuclear Transfer
In SCNT, an egg cell is taken from an animal and its nucleus is removed, creating an enucleated egg. Then a somatic cell—any cell that is not a sex...
Cloning of Dolly the Sheep
The first successfully cloned mammal was Dolly, a sheep, born on 5th July 1996 at Roslin Institute, Scotland. The cloned sheep was named after the American singer Dolly Parton. Dolly lived for seven years and died of respiratory complications, which is speculated to be due to the actual age of her DNA. Because the DNA in cloned cells belongs to an older individual, the cloned individual’s life expectancy may be affected. Indeed, analysis of Dolly’s DNA revealed shorter telomeres than other...


