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J Biomol Tech. 2010 September; 21(3 Suppl): S48–S49.
PMCID: PMC2918093

Influenza A H1N1: A Rapid Approach to Cloning and Protein Expression

K. Kwon,1 G. Tsegaye,1 P. Burr,1 Y. Do,1 J. Sosa,1 T. Edeto,1 M. Gonzalez,1 P. Tangirala,1 B. Bishop,1 R. Sanka,1 J. Gilbert,1 M. Urh,2 J. Vidugiriene,2 D. Hartzell,2 R. Fleischmann,1 and S. Peters1
1J. Craig Venter Institute, Rockville, MD, United States;
2Promega Corporation, Madison, WI, United States

Abstract

RP-88

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/CDSs 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.


Articles from Journal of Biomolecular Techniques : JBT are provided here courtesy of The Association of Biomolecular Resource Facilities