tailieunhanh - báo cáo hóa học:" Mutational study of sapovirus expression in insect cells"

Tuyển tập các báo cáo nghiên cứu về hóa học được đăng trên tạp chí sinh học đề tài : Mutational study of sapovirus expression in insect cells | Virology Journal BioMed Central Research Open Access Mutational study of sapovirus expression in insect cells Grant S Hansman Kazuhiko Katayama Tomoichiro Oka Katsuro Natori and Naokazu Takeda Address Department of Virology II National Institute of Infectious Diseases Tokyo Japan Email Grant S Hansman - ghansman@ Kazuhiko Katayama - katayama@ Tomoichiro Oka - oka-t@ Katsuro Natori - natori@ Naokazu Takeda - ntakeda@ Corresponding author Published 23 February 2005 Received 28 January 2005 Accepted 23 February 2005 Virology Journal 2005 2 13 doi 1743-422X-2-13 This article is available from http content 2 1 13 2005 Hansman et al licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License http licenses by which permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited. Abstract Human sapovirus SaV an agent of human gastroenteritis cannot be grown in cell culture but expression of the recombinant capsid protein rVP1 in a baculovirus expression system results in the formation of virus-like particles VLPs . In this study we compared the time-course expression of two different SaV rVP1 constructs. One construct had the native sequence Wt construct whereas the other had two nucleotide point mutations in which one mutation caused an amino acid substitution and one was silent MEG-1076 construct . While both constructs formed VLPs morphologically similar to native SaV Northern blot analysis indicated that the MEG-1076 rVP1 mRNA had increased steady-state levels. Furthermore Western blot analysis and an antigen enzyme-linked immunosorbent assay showed that the MEG-1076 construct had increased expression levels of rVP1 and yields of VLPs. Interestingly the position of the mutated residue was strictly conserved residue among other human SaV strains suggesting