Ubiquitin-fusion protein system: a powerful tool for ectopic protein expression in mammalian cells
نویسندگان
چکیده
منابع مشابه
Ubiquitin-fusion protein system: a powerful tool for ectopic protein expression in mammalian cells.
Clonal cell lines stably expressing a protein of interest are frequently used in the characterization of the potential physiological functions of mammalian proteins, but the generation of such cell lines is laborious and time-consuming. Here, we made use of the fact that fusion proteins consisting of ubiquitin linked to the N terminus of a protein of interest are efficiently processed by ubiqui...
متن کاملIMPROVEMENT OF hGM-CSF EXPRESSION USING A FUSION SYSTEM BY DIRECTING PROTEIN TO THE PERIPLASMIC SPACE
Bacterial expression systems provide economic and logistic advantages in manufacturing proteins for human therapeutic purposes. However, most such proteins accumulate in insoluble biologicaIIy inactive form when overexpressed in bacterial cells. This is the case while attempting to produce recombinant human granulocyte-macrophage colony stimulating factor (hGM-CSF) in E. coli. In this repor...
متن کاملSplit-Ubiquitin Based Membrane Yeast Two-Hybrid (MYTH) System: A Powerful Tool For Identifying Protein-Protein Interactions
The fundamental biological and clinical importance of integral membrane proteins prompted the development of a yeast-based system for the high-throughput identification of protein-protein interactions (PPI) for full-length transmembrane proteins. To this end, our lab developed the split-ubiquitin based Membrane Yeast Two-Hybrid (MYTH) system. This technology allows for the sensitive detection o...
متن کاملMycobacterium tuberculosis HspX/EsxS Fusion Protein: Gene Cloning, Protein Expression, and Purification in Escherichia coli
Background: The purpose of this study was to clone, express, and purify a novel multidomain fusion protein of Micobacterium tuberculosis (Mtb) in a prokaryotic system. Methods: An hspX/esxS gene construct was synthesized and ligated into a pGH plasmid, E. coli TOP10 cells were transformed, and the vector was purified. The vector containing the construct and pET-21b (+) plasmid were digested ...
متن کاملCloning and Expression of Simian Rotavirus Spike Protein (VP4) in Insect Cells by Baculovirus Expression System
Background: VP4 protein is as spikes on rotavirus outer capsid shell which is responsible for virus attachment to the host. VP4 induces production of neutralizing antibodies which could be used for serotyping of different isolates. Methods: Simian rotavirus SA11 gene 4 cDNA was cloned into a cloning plasmid pDONRTM by recombination reaction using clonase II enzyme mix. The resulting clone was c...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
ژورنال
عنوان ژورنال: BioTechniques
سال: 2009
ISSN: 0736-6205,1940-9818
DOI: 10.2144/000113023