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 Peterson^1
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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