National Institute of Virology, 20-A, Dr Ambedkar Road, Pune-411 011, India.
Abstracts of the papers presented at the XVIII National Conference of Indian Virological Society at Post Graduate Institute of Medical Education and Research, Chandigarh, India, December 11–13, 2008.
Large-scale epidemic caused by Chikungunya virus leads to high morbidity, affecting millions of people. It has been reemerged in India in 2005 and is ongoing still in southern part of India in 2008. Chikungunya envelope proteins (E1 and E2) are responsible for the life long immunity against the CHIK virus. The genes coding for these proteins were amplified by PCR from the cDNA of 26S RNA coding for all structural proteins of chikungunya and cloned in bacterial expression vector Pet32b+ at BamHI/HindIII restriction sites downstream to the gene of thioredoxin (Trx) and histidine 6aa.tag to give the pET-32b/CHIKE1 and the pET-2b/CHIKE2 plasmids. The recombinant proteins E1-E2 expressed in inclusion bodies (IBs) and were purified using BugBuster protein reagent and protein refolding kit (Novagen). Two recombinant proteins were affinity-purified by Ni-charged His-binding resin (Novagen). These proteins were immune-reactive against sera from rabbit, mouse and guinea pig previously immunized with native CHIK virus, which confirms the antigencity of recombinant E1 and E2 proteins expressed in the Escherichia coli system.
