Construction Of Rotavirus VP4 Protein To The RRS Screening System Vector

The yeast two-hybrid system has been one of the important methods for research of interaction between proteins since it was founded. Traditional two-hybrid locates the interaction in the nucleus, while the Ras recruitment system (RRS) developed in recent years can locate that on the cell membrane and has more advantages such as high sensitivity and less false positive. In our study, we verified the RRS screening system through interaction between two known proteins, for the purpose of applying this system for wider research on the interaction between other proteins and obtaining the interacted proteins.RV gastroenteritis and its complications threaten the health of children seriously. The first and necessary step for RV infection is its binding to the receptors on the cell membrane. Therefore, it is so important to identify the nature of these receptors for better precaution and treatment of those diseases. The VP4 spike protein of RV is the virus attachment protein binding to the receptor. In our study, the cDNA fragment of VP4 protein was amplified by RT-PCR and cloned in the plasmid Met425-Myc-Ras to obtain a bait plasmid suitable for RRS screening system, and the self-activation of this bait was excluded. The above fact showed that we can employ this bait and the RRS screening system to draw the RV receptors from the cDNA library of RV susceptible cell.