재조합 아데노 바이러스 생산을 위한 효과적인 Shuttle 벡터의 개발과 검증

Abstract

Recombinant adenovirus is an attractive vehicle to deliver a foreign gene for the virus can infect many cell types including non-dividing cells. Production of a recombinant adenovirus requires co-transfection of a shuttle vector which harbors a gene of interest and a rescue vector. Adenoviral shuttle vectors currently in use contain limited numbers of cloning sites for insertion of a foreign gene. They also lack a promoter and a signaling sequence for the poly(A) tail. Therefore, we set out to construct two adenoviral shuttle vectors, pAd-YC and pAd-YC2, in an effort to facilitate cloning and expression of foreign genes. pAd-YC contains the CMV promoter and multiple cloning sites but without a poly(A) signal sequence. pAd-YC2 contains the CMV promoter, the multiple cloning sites and the poly(A) signal sequence. The functionality of pAd-YC and pAd-YC2 was tested by cloning and expression of a chimeric Fcγreceptor cDNA and the β-galactosidase gene. Fcγreceptor expression was confirmed by binding of opsonized erythrocytes to COS-1 infectants, and β-galactosidase expression was assayed for X-gal staining. The results demonstrate the functional efficacy of the two shuttle vectors. We have since successfully constructed several recombinant adenoviruses by employing the two versatile shuttle vectors.