Preparation method of recombinant Fas-associated death domain protein and application thereof

Abstract

The invention belongs to the technical field of biology, in particular to a preparation method of full-length Fas-associated death domain (FADD) protein, which utilizes a recombinant DNA technique to express and prepare the full-length FADD protein which has good stability, high yield, the same high-level structure as a wild FADD and activity for inducing cell apoptosis, and the preparation method has more convenient and faster separation and purification process and high yield. The full-length FADD protein can be applied to prepare a medicament for inducing the cell apoptosis and treat diseases such as tumors, rheumatoid arthritis, and the like. The invention also provides an expression method for expressing the full-length FADD protein in a prokaryotic expression system, which can effectively improve the yield of the full-length FADD protein.

Description

1. Technical field: [0001] The invention belongs to the field of genetic engineering biotechnology. 2. Background technology: [0002] FADD (Fas-associated death domain protein, Fas-associated death domain protein) is a protein that can specifically interact with the cytoplasmic region of the death receptor Fas, which was screened by two research groups such as Chinnaiyan and Boldin in 1995 using the yeast two-hybrid system. The protein is named as Fas-associated death domain protein (Fas-associated death domain protein, FADD) because it contains a highly homologous domain with the death domain of Fas protein [Boldin, M.P. et al., 1995, J. Biol. Chem. 270:7795-7798; Chinnaiyan, A.M. et al., 1995, Cell 81:505-512]. The human FADD gene encodes 208 amino acids with a molecular weight of 23kDa, which can be divided into three functional regions, including DED (death effect domain, death effect domain) of about 76 amino acids at the N-terminal and 70 amino acids at the C-termina...

AI Technical Summary

Problems solved by technology

It can be seen that the above FADDs are all fragments of FADD, among which the FADD of amino acid residues 1-191 expressed by Carrington P.E. et al. in 2006 is the longest, but it still lacks 17 amino acid residues at positions 192-208

Although in the above studies, scientists have tried various methods to obtain the various truncated FADD proteins for partial functional and structural studies, currently no laboratory can express and obtain full-length FADD proteins in large quantities. The reason is that FADD protein is unstable and very easy to aggregate, so even if a higher level of FADD protein expression is obtained by using genetic engineering technology, it is impossible to separate and purify a sufficient amount of FADD protein for research or even treatment.

Images