Cabrita Lisa D, Bottomley Stephen P
Monash University, Department of Biochemistry and Molecular Biology, School of Biomedical Sciences, P.O. Box 13D, Melbourne, Victoria 3800, Australia.
Biotechnol Annu Rev. 2004;10:31-50. doi: 10.1016/S1387-2656(04)10002-1.
The release of sequence data, particularly from a number of medically and biotechnologically important genomes, is increasing in an exponential fashion. In light of this, elucidating the structure and function of proteins, particularly in a "high throughput" manner, is an important quest. The production of recombinant proteins however is not always straightforward, with a number of proteins falling prey to low expression problems, a high susceptibility to proteolysis and the often despised production of inclusion bodies. Whilst expression as inclusion bodies can often be advantageous, their solubilization and renaturation is often a time consuming and empirical process. In this review, we aim to outline some of the more common approaches that have been applied to a variety of proteins and address issues associated with their handling.
序列数据的发布,尤其是来自许多医学和生物技术领域重要基因组的序列数据,正呈指数级增长。鉴于此,阐明蛋白质的结构和功能,尤其是以“高通量”方式进行阐明,是一项重要的探索。然而,重组蛋白的生产并非总是一帆风顺,许多蛋白质存在表达量低、易被蛋白酶水解以及经常令人头疼的包涵体形成等问题。虽然以包涵体形式表达通常可能具有优势,但其溶解和复性往往是一个耗时且凭经验的过程。在本综述中,我们旨在概述一些已应用于多种蛋白质的较为常见的方法,并探讨与其处理相关的问题。