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Báo cáo hóa học: " Recombination-ready Sindbis replicon expression vectors for transgene expression"

Tham khảo tài liệu 'báo cáo hóa học: " recombination-ready sindbis replicon expression vectors for transgene expression"', luận văn - báo cáo phục vụ nhu cầu học tập, nghiên cứu và làm việc hiệu quả | Virology Journal BioMed Central Methodology Open Access Recombination-ready Sindbis replicon expression vectors for transgene expression Brian J Geiss Lisa H Shimonkevitz Cherilyn I Sackal and Ken E Olson Address Arthropod-Borne and Infectious Diseases Laboratory Department of Molecular Biology. Immunology and Pathology Colorado State University Fort Collins CO 80523 USA Email Brian J Geiss - Brian.Geiss@colostate.edu Lisa H Shimonkevitz - Lisa.Shimonkevitz@colostate.edu Cherilyn I Sackal - Cherilyn.Sackal@colostate.edu Ken E Olson - Kenneth.Olson@colostate.edu Corresponding author Published 26 October 2007 Received 18 September 2007 Accepted 26 October 2007 Virology Journal 2007 4 112 doi 10.1186 I743-422X-4-II2 This article is available from http www.virologyj.com content 4 1 1 12 2007 Geiss et al licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License http creativecommons.org licenses by 2.0 which permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited. Abstract Background Sindbis viruses have been widely used as tools to study gene function in cells. Despite the utility of these systems the construction and production of alphavirus replicons is time consuming and inefficient due to potential additional restriction sites within the insert region and lack of directionality for insert ligation. In this report we present a system useful for producing recombinant Sindbis replicons that uses lambda phage recombination technology to rapidly and specifically construct replicon expression plasmids that contain insert regions in the desired orientation. Results Recombination of the gene of interest with the replicon plasmid resulted in nearly 100 recombinants each of which contained a correctly orientated insert. Replicons were easily produced in cell culture and packaged into pseudo-infectious viral particles. Insect and mammalian cells .