Influenza A H1N1: A Rapid Approach to Cloning and Protein Expression
Part # PS103
Abstract
Jeremy Hasseman1, Keehwan Kwon1, Getahun Tsegaye1, Patrick Burr1, Yu Do1, Julia Sosa1, Tigist Edeto1, Monica Gonzalez1, Padmavathi Tangirala1, Jason Stam1 Sarah Grimshaw1, Carissa Grose1, Brian Bishop1, Ravi Sanka1, Marjeta Urh2, Nidhi Nath2, Jolanta Vidugiriene2, Danette Hartzell2, Robert Fleischmann1, Scott Peterson1
1Pathogen Functional Genomics Resource Center, J. Craig Venter Institute, Rockville, MD
2Promega Corporation, Madison, WI
In May 2009, Swine-derived Influenza A H1N1 virus emerged as a serious threat to global health. With the threat came the need for the virology community to rapidly characterize this emergent subtype. To enable efficient distribution of materials for further characterization, a rapid cloning and protein expression process was implemented using two clinical isolates. Clones were made of all genomic segments/protein coding regions using three different cloning systems. Proteins were expressed in a cell-free system with a HaloTag™ fusion and immobilized on a HaloLink™ microarray substrate. The immobilized proteins were then challenged with anti-H1N1 antibodies to assess immunosensitivity. Mammalian and baculoviral (BEVS) systems were utilized to ensure glycosylation of HA and NA proteins. Clones were publicly available and protein microarrays made within one month after receiving H1N1 DNA.
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