【交流】核酸版2005年7月份明星技术: In vitro site-specific mutagenesis
丁香园论坛
661
In vitro site-specific mutagenesis
An in vitro technique in which an alteration is made at a specific site in a DNA molecule, which is then reintroduced into a cell.Various techniques are used; for the cell biologist,a very powerful approach to determining which parts of a protein or nucleotide sequence is critical to function.
Site specific mutagenesis was invented in 1978 by Clyde Hutchison and Michael Smith. Michael Smith was given the Nobel Prize for discovering it in 1993.
The mutagenesis can be conducted in vivo (in studies of model organisms, or cultured cells) or in vitro and the mutagenesis can be directed to a specific site in a pre-determined way (site-directed mutagenesis), or can be random. In the case of in vivo mutagenesis, for example, gene targeting offers exquisite site-directed mutagenesis within living cells while exposure of male mice to high levels of a powerful mutagen such as ethyl nitrosurea (ENU) and subsequent mating of the mice offers a form of random mutagenesis which can be important in generating new mutants .
In vitro mutagenesis can involve essentially random approaches to mutagenesis, which may be valuable in producing libraries of new mutants. In addition, if a gene has been cloned and a functional assay of the product is available, it is also very useful to be able to employ a form of in vitro mutagenesis, which results in alteration of a specific amino acid or small component of the gene product in a predetermined way.
加分原则：
1. 你可以在此跟帖提问。如果你的问题有一定深度，引发战友们积极讨论和思考。加1-2分。
2. 回答问题或介绍经验。可加1-2分。
3. 精品论文导读（Cell,Nature,Science,Nature Genetics,PNAS等杂志上的更易加分），可加1-3分，即给出全文链接，且深入浅出介绍文章的主要内容加1分；且对文章的实验设计、结果和本文可能开展后续工作的介绍加2分；同时用流线图画出文章的技术路线、思路或结构的加3分。
站友们在发知识性的帖子时，请注意标明“原创”或“转帖（及其来源）”。因为原创加分力度会大一点。多谢合作！
本版前期的明星技术：
1. 【交流】核酸版2005年6月份明星技术: nuclear run on transcription
2. 【交流】核酸版2005年5月份明星技术: Switching Mechanism At 5' end of the RNA Transcript: SMART
3. 【交流】核酸版2005年4月份明星技术： Electrophoretic Mobility Shift Assay, EMSA
4. 【交流】核酸版2005年3月份明星技术：Chromatin immunoprecitation,ChIP
5. 【交流】核酸版2005年02月份明星技术：SELEX技术
6. 【交流】核酸版2005年元月份明星技术：Ribonuclease protection assay，RPA
7. 【交流】核酸版2004年12月份明星技术：Serial Analysis of Gene Expression，SAGE
8. 【交流】核酸版2004年10月份明星技术：DNA Microarray
An in vitro technique in which an alteration is made at a specific site in a DNA molecule, which is then reintroduced into a cell.Various techniques are used; for the cell biologist,a very powerful approach to determining which parts of a protein or nucleotide sequence is critical to function.
Site specific mutagenesis was invented in 1978 by Clyde Hutchison and Michael Smith. Michael Smith was given the Nobel Prize for discovering it in 1993.
The mutagenesis can be conducted in vivo (in studies of model organisms, or cultured cells) or in vitro and the mutagenesis can be directed to a specific site in a pre-determined way (site-directed mutagenesis), or can be random. In the case of in vivo mutagenesis, for example, gene targeting offers exquisite site-directed mutagenesis within living cells while exposure of male mice to high levels of a powerful mutagen such as ethyl nitrosurea (ENU) and subsequent mating of the mice offers a form of random mutagenesis which can be important in generating new mutants .
In vitro mutagenesis can involve essentially random approaches to mutagenesis, which may be valuable in producing libraries of new mutants. In addition, if a gene has been cloned and a functional assay of the product is available, it is also very useful to be able to employ a form of in vitro mutagenesis, which results in alteration of a specific amino acid or small component of the gene product in a predetermined way.
加分原则：
1. 你可以在此跟帖提问。如果你的问题有一定深度，引发战友们积极讨论和思考。加1-2分。
2. 回答问题或介绍经验。可加1-2分。
3. 精品论文导读（Cell,Nature,Science,Nature Genetics,PNAS等杂志上的更易加分），可加1-3分，即给出全文链接，且深入浅出介绍文章的主要内容加1分；且对文章的实验设计、结果和本文可能开展后续工作的介绍加2分；同时用流线图画出文章的技术路线、思路或结构的加3分。
站友们在发知识性的帖子时，请注意标明“原创”或“转帖（及其来源）”。因为原创加分力度会大一点。多谢合作！
本版前期的明星技术：
1. 【交流】核酸版2005年6月份明星技术: nuclear run on transcription
2. 【交流】核酸版2005年5月份明星技术: Switching Mechanism At 5' end of the RNA Transcript: SMART
3. 【交流】核酸版2005年4月份明星技术： Electrophoretic Mobility Shift Assay, EMSA
4. 【交流】核酸版2005年3月份明星技术：Chromatin immunoprecitation,ChIP
5. 【交流】核酸版2005年02月份明星技术：SELEX技术
6. 【交流】核酸版2005年元月份明星技术：Ribonuclease protection assay，RPA
7. 【交流】核酸版2004年12月份明星技术：Serial Analysis of Gene Expression，SAGE
8. 【交流】核酸版2004年10月份明星技术：DNA Microarray
