Cloning, Expression and Purification of Haemagglutinin and Neuraminidase gene of highly Pathogenic Avian Influenza H5N1 in Escherichia coli

Order of Publishing in Issue: 
10
Volume :6
Issue :2
April, 2012
Page No: 
222-228
Authors: 
M. Subathra [1], P. Santhakumar [2], P. Pardhasaradhi [1] , M. Lakshmi Narasu [1]*, Chandrani Chakravarty3 and Sunil K. Lal3
Address: 
[1] Centre for Biotechnology, Jawaharlal Nehru Technological University, Hyderabad, India
Address: 
[2] Indian Immunologicals Ltd, Hyderabad, India
Address: 
[3] Virology Group, International Centre for Genetic Engineering and Biotechnology (ICGEB), New Delhi, India
Email-ID: 
mangamoori@gmail.com

Abstract
The looming influenza virus pandemic requires simple and quicker vaccination strategies to prevent higher mortality and morbidity both in chickens and in humans. The process of current influenza vaccines manufacturing using embryonated eggs cannot help in controlling a future pandemic as it is too slow and the yield obtained by these methods are quit lower. In this study, we have developed a bacterial expressed rHA and rNA for using as a subunit vaccine against avian influenza which could also be used as a diagnostic tools against avian influenza. The HA and NA genes were cloned individually into Escherichia coli expression vector pRSETA. The rHA and rNA were expressed in E.coli and were purified using Ni-NTA chromatography. The haemagglutinin activity of the E.coli expressed rHA was analyzed against various RBCs. The rHA and rNA expressed E.coli can be used as a quicker and cheaper subunit vaccine candidate against avian influenza and also as a tools for diagnosis.

Keywords: 
Highly pathogenic avian influenza, haemagglutinin, neuraminidase, vaccine, diagnosis
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