School of Chemical Sciences, The University of Auckland, Private Bag 92019, Victoria Street West, Auckland, 1142, New Zealand.
Facultad de Ingeniería, Universidad Nacional Autónoma de México, Av. Universidad 3000, Ciudad Universitaria, Coyoacán, Cd. Mx., CP 04510, Mexico.
Sci Rep. 2019 Jun 20;9(1):8884. doi: 10.1038/s41598-019-45323-8.
Human ribosomal proteins play important structural and functional roles in the ribosome and in protein synthesis. An efficient method to recombinantly produce and purify these proteins would enable their full characterisation. However, the production of human ribosomal proteins can be challenging. The only published method about the recombinant production of human ribosomal proteins involved the recovery of proteins from inclusion bodies, a process that is tedious and may lead to significant loss of yield. Herein, we explored the use of different Escherichia coli competent cells and fusion protein tags for the recombinant production of human ribosomal proteins. We found that, by using thioredoxin as a fusion protein, soluble ribosomal protein could be obtained directly from cell lysates, thus leading to an improved method to recombinantly produce these proteins.
人类核糖体蛋白在核糖体和蛋白质合成中发挥着重要的结构和功能作用。一种高效的重组生产和纯化这些蛋白质的方法将能够对其进行全面的表征。然而，生产人类核糖体蛋白可能具有挑战性。关于重组生产人类核糖体蛋白的唯一已发表方法涉及从包涵体中回收蛋白质，这是一个繁琐的过程，可能会导致产量显著损失。在此，我们探索了使用不同的大肠杆菌感受态细胞和融合蛋白标签来重组生产人类核糖体蛋白。我们发现，通过使用硫氧还蛋白作为融合蛋白，可以直接从细胞裂解物中获得可溶性核糖体蛋白，从而为重组生产这些蛋白质提供了一种改进的方法。
