Shatzman A R, Rosenberg M
Hepatology. 1987 Jan-Feb;7(1 Suppl):30S-35S. doi: 10.1002/hep.1840070706.
There are numerous proteins of biological interest which cannot be obtained from natural sources in quantities sufficient for detailed biochemical and physical analysis. The limited bioavailability of these molecules has made it impossible to consider their potential utilization as either pharmacological agents and/or targets. One solution to this problem has been the development of recombinant vector systems which are designed to achieve efficient expression of cloned genes in a variety of biological systems. This paper will describe the development and application of a particular set of vectors which have been designed to achieve efficient expression of essentially any gene coding sequence in Escherichia coli. The system utilizes efficient phage-derived transcriptional and translational regulatory signals and provides a strong regulatable promoter, an antitermination mechanism to ensure efficient transcription across any gene insert, high stability and, when appropriate, efficient translation initiation information. In addition, a wide variety of host strains have been developed in order to help control, stabilize and maximize expression of various cloned genes. The system has now been used to express efficiently more than 75 different prokaryotic and eukaryotic gene products. The application of this system to the expression and characterization of several oncogene products will be described.
有许多具有生物学意义的蛋白质无法从天然来源获得足够数量以进行详细的生化和物理分析。这些分子有限的生物可利用性使得无法考虑将其作为药物和/或靶点进行潜在利用。解决这个问题的一种方法是开发重组载体系统，该系统旨在在多种生物系统中实现克隆基因的高效表达。本文将描述一组特定载体的开发和应用，这些载体旨在在大肠杆菌中实现基本上任何基因编码序列的高效表达。该系统利用高效的噬菌体衍生转录和翻译调控信号，并提供一个强可调控启动子、一种抗终止机制以确保跨任何基因插入片段的高效转录、高稳定性以及在适当情况下高效的翻译起始信息。此外，还开发了多种宿主菌株，以帮助控制、稳定和最大化各种克隆基因的表达。该系统现已用于高效表达75种以上不同的原核和真核基因产物。将描述该系统在几种癌基因产物的表达和表征中的应用。
