IMMUNO INFORMATIC APPROACHES IN DESIGNING VACCINE AGAINST PATHOGENIC LEPTOSPIRA THROUGH PAN GENOME REVERSE VACCINOLOGY
| dc.contributor.advisor | RANIPRAMEELA, D(MAJOR) | |
| dc.contributor.advisor | VINOD KUMAR, N | |
| dc.contributor.advisor | JAGADEESH BABU, A | |
| dc.contributor.author | SUDHEER, P | |
| dc.date.accessioned | 2018-11-30T12:03:01Z | |
| dc.date.available | 2018-11-30T12:03:01Z | |
| dc.date.issued | 2016-12 | |
| dc.description | THESES | en_US |
| dc.description.abstract | ABSTRACT: Leptospirosis is a globally important zoonotic disease caused by pathogenic Leptospira and it is a disease of livestock, pet animals, wildlife and humans throughout the world. The losses are due to reproductive problems in livestock and mortality in case of humans. Current existing leptospiral vaccines are unsuccessful due to their limitations .Hence there is a need to develop a novel and efficacious vaccine to control the disease. To overcome this, reverse vaccinology is the right choice. In recent trends, it is possible to target the common vaccine candidates with genomic information of the single organism. Based on this concept the present work was planned to study the Bioinformatics approaches to identify the vaccine candidates in designing a vaccine against pathogenic Leptopsira in Bovines In the present study complete proteomes of L.borgpetersenii hardjo bovis JB 197 and L550 were screened to identify common surface exposed proteins through Rxvi language scripts and codes. Later these common surface exposed proteins were subjected to DEG analysis. 49 essential proteins were identified .Further essential proteins were subjected to non-homology analysis against both host and gut microbiota to avoid autoimmunity using NCBI-BLAST.T-helper cell epitopes were predicted from non-homologous proteins through MetaMHCII and ProPred homology search against BoLA-DRB3, and evaluated using Vaxijen server. Three dimensional structures were built for T-cell epitopes and BoLA-DRB3 using Modeller9v.15. A total of twenty five models were generated. The model with high negative DOPE score was selected and validated using PROCHECK, ProQ, and ProSA in determining protein quality. The structures of T-cell epitopes and BoLA DRB3 were prepared before docking using protein preparation wizard of Schrodinger 2015-3. Docking and free energy calculations were performed with BioLuminate Module v 2.0 of Schrödinger software suite 2015-3. The changes in structural confirmation were monitored in terms of energy plot, RMSD and RMSF during 50 ns MD simulations run time using Desmond v4.3. In Silico analysis of L.borgpetersenii JB 197 and L550 retrieved three proteins namely Ton B dependent receptor containing single epitope, ABC permease protein with three epitopes and UVr ABC protein B with single epitope. L.ballum was selected instead of L.borgpetersenii due to its non-availability of the culture during the period of the study and 99% identity on BLASTp. The nucleotide sequence corresponding to ABC permease gene containing three epitopes was retrieved, primers were designed and PCR was standardized for the amplification of ABC permease gene. PCR purified product on sequencing analysis confirmed the presence of ABC permease gene. Then, the PCR purified product was cloned in to PRSET vector using E.coli DH5α cells .The recombinant plasmid was transformed in to E.coli BL21 (DE3) cells and expression was induced by addition of 1mM IPTG. Finally recombinant protein was extracted from lysate of E.coli BL21 (DE3) cells. Recombinant protein was analysed on SDS-PAGE for characterization. The SDS-PAGE analysis yielded 20KD of expected recombinant protein on staining with commassie brilliant blue | en_US |
| dc.identifier.uri | http://krishikosh.egranth.ac.in/handle/1/5810084926 | |
| dc.keywords | IMMUNO INFORMATIC ;DESIGNING VACCINE;PATHOGENIC LEPTOSPIRA;PAN GENOME;REVERSE VACCINOLOGY | en_US |
| dc.language.iso | en | en_US |
| dc.pages | 140 | en_US |
| dc.publisher | SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA | en_US |
| dc.sub | Veterinary Microbiology | en_US |
| dc.subject | null | en_US |
| dc.theme | IMMUNO INFORMATIC APPROACHES IN DESIGNING VACCINE AGAINST PATHOGENIC LEPTOSPIRA THROUGH PAN GENOME REVERSE VACCINOLOGY | en_US |
| dc.these.type | M.V.Sc. | en_US |
| dc.title | IMMUNO INFORMATIC APPROACHES IN DESIGNING VACCINE AGAINST PATHOGENIC LEPTOSPIRA THROUGH PAN GENOME REVERSE VACCINOLOGY | en_US |
| dc.type | Thesis | en_US |