“EXPRESSION, PURIFICATION AND QUANTIFICATION OF NEURAMINIDASE GENE OF INFLUENZA” A dissertation submitted to the VIT University in partial fulfillment of the requirement for the award of the degree of
MASTER OF SCIENCE In BIOTECHNOLOGY SUBMITTED BY A. SRIKANTH
Register No: 08MSBOO1
(Estd. u/s 3 of UGC Act 1956)
SCHOOL OF BIO SCIENCES AND TECHNOLOGY VIT UNIVERSITY VELLORE-632014 MAY 2010
I here by declare that the project work entitled “EXPRESSION,PURIFICATION AND CHARACTERIZATION OF NEURAMINIDASE GENE OF INFLUENZA” submitted in partial fulfillment of the requirement for the Master of Science in Biotechnology from School of Biotechnology Chemical and Biomedical Engineering, VIT UNIVERSITY. The project is the result of work carried out under the supervision of Dr. Archana Giri, Head, CBT, IST, and JNTU-HYDERABAD.
Avian Influenza outbreaks among poultries are occur world wide from time to time with increasing Global pandemics. The strategies to prevent and test for avian influenza are becoming increasingly more important. One strain of avian influenza, H5N1, has already crossed the species border into humans. We propose a method for developing a subunit vaccine and influenza tests using Bacteria as the chassis, respectively. This system is easily amendable for targeting the two main antigens presented on the exterior of the influenza virus: Hemagglutinin and Neuraminidase. The purification and influenza vaccine & Diagnostics development. Recombinant envelop protein of H5N1, 55KDa of rNA1, was successfully produced in prokaryotic system by using pRSET/A expression vector, and the expression was optimized by IPTG, this shows the ideal condition to obtain high yield of rNA1 protein. The result of purification showed that the expressed protein purified form of rNA1.The quantification analysis through calorimetric assay revealed yield of recombinant protein rNA1 protein respectively. This experimental procedure completely shows production of recombinant AIV/NA protein in bacterial chassis especially in E coli for high yields at low cost & lesser time effectively. quantification Of Neuraminidase protein in this study will provide useful information further avian
ACKNOWLEDGEMENT Apart from the efforts of me, the success of this project depends largely on encouragement and guidelines of many others. I take this opportunity to express my gratitude to the people who have been instrumental in the successful completion this project With a deep sense of gratitude, I record my sincere thanks to Dr. Archana Giri, Head, CBT, IST, JNTUH, HYDERABAD, for providing exhaustive facilities and Government funded project in Biotech Division. Her constant encouragement and valuable guidance and support have helped me to complete the work on time. I record my sincere thanks to Shri. G. Viswanathan, Chancellor, VIT University, Vellore. I am thankful to Dr. Lazer Mathew, Dean, SBST -VIT University for his support and encouragement in this project. I extend my sincere thanks to Division leader Dr Murali Manoj and other teaching and non teaching members of biotech Division, SBST, VIT University for helping us all the way. I would like to thank Dr .C.Shanthi, Professor, VIT University for her valuable suggestions as an internal guide, and my sincere thanks to other teaching and non teaching members of biotech Division, SBST, VIT University for helping us all the way. With immense pleasure and respect I express my deep sense of gratitude to Mr. Jyotheswar Reddy (All India best PhD scholar awarder by European Union) for his constant encouragement and inspiration throughout this period to complete the project successfully. Special thanks to Mr. Shaik Abdullah, Miss Monica Reddy for their constant help on course of this study. Finally the, love, understanding and constant encouragement from my family is always remembered. The usefulness of this thesis, I dedicated to...
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