Molecular studies on the toxifying effects by genetically engineered cytochromes P450.
Molecular studies on the toxifying effects by genetically engineered cytochromes P450.
复制标题
基因工程细胞色素 P450 毒性作用的分子研究。
DOI：
10.1081/dmr-100101928
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发表时间：
1999
影响因子：
5.9
通讯作者：
Greim,H
中科院分区：
文献类型：
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作者：
Doehmer,J;Buters,JT;Luch,A;Soballa,V;Baird,WM;Morisson,H;Stegeman,JJ;Townsend,AJ;Greenlee,WF;Glatt,HR;Seidel,A;Jacob,J;Greim,H
经过近二十年的发展，现在很明显，基于分子生物学和基因技术的方法学已经极大地改变了基础和应用毒理学的方式。它始于1980年，通过鉴定和克隆编码参与异生物质代谢的细胞色素P450同工酶的基因，并继续在合适的宿主如细菌、酵母、昆虫和哺乳动物细胞中表达来自不同物种（包括人）的克隆基因[1]。这些cDNA衍生的表达系统与其他基因工程工具[例如，聚合酶链反应（PCR）和先进的合成和分析化学方法]相结合，形成了一个新的技术平台，用于研究代谢依赖性毒性作用，其详细程度超过了更经典的方法（例如，动物研究）。这些现代化的工具是精确的，具体的，在毒理学的许多应用程序的通用性。这取决于宿主细胞的生物学和遗传结构，这些工具最适合研究毒理学问题。我们已经给了优势，以基因工程的中国仓鼠细胞的细胞系V79。自20世纪50年代建立以来，该细胞系在毒理学测试中具有悠久的传统[2]。它是一个加倍速度非常快的细胞系，其加倍速度小于12 h，具有二倍体和稳定的核型，具有一条X染色体，其克隆效率高于90%。因此，我们认为，
After almost two decades, it is now evident that methodology based on molecular biology and gene technology has dramatically changed the way basic and applied toxicology is being performed. It started in 1980 by identifying and cloning genes encoding cytochromes P450 isoenzymes involved in xenobiotic metabolism and has been continued with the expression of cloned genes from different species, including man, in suitable hosts such as bacterial, yeast, insect, and mammalian cells [1]. These cDNA-derived expression systems in combination with other genetically engineered tools [eg, polymerase chain reaction (PCR) and advanced synthetic and analytical chemical methods] form a new technology platform for studying metabolism-dependent toxic effects in greater detail than it is possible by a more classical approach (eg, animal studies). These modern tools are precise, specific, and versatile for many applications in toxicology. It depends on the biology and the genetic construction of the host cell, which these tools are best suited to investigate a toxicological problem. We have given advantage to genetically engineered Chinese hamster cells of the cell line V79. This cell line has a long-standing tradition in toxicological testing since its establishment in the 1950’s [2]. It is a cell line with an unusually fast doubling rate of less than 12 h, it has a diploid and stable karyotype with one X chromosome, and its cloning efficiency is higher than 90%. Therefore,