Recombination activating gene (RAG)-1 and 2 encoding proteins expressed by the baculovirus system

Araki R, Kaku M, Itoh M, Hamatani K, Usui T, Abe M
Acta Med Nagasaki 41(1):43-7, 1996

Summary

We have been attempting to obtain mouse recombination activating gene-1 (RAG-1) and RAG-2 protein for biochemical analyses. First of all, we obtained truncated products of these genes expressed and purified using the E. coli expression system and then established the polyclonal and monoclonal antibodies by means of E. coli expressed peptides as antigens. Subsequently, whole RAG-1 and RAG-2 gene products were expressed the baculovirus expression system. Since it has been difficult to achieve the significant gene expression of full-length cDNA, we employed the glutathione S-transferase (GST)-fused gene-expression system which facilitated the massive expression of gene products. This system was also advantageous in that we could detect the expressed protein molecules not only with anti-RAG antibody but also with anti-GST antibody.