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University of Agricultural Sciences, Bengaluru

University of Agricultural Sciences Bangalore, a premier institution of agricultural education and research in the country, began as a small agricultural research farm in 1899 on 30 acres of land donated by Her Excellency Maharani Kempa Nanjammanni Vani Vilasa Sannidhiyavaru, the Regent of Mysore and appointed Dr. Lehmann, German Scientist to initiate research on soil crop response with a Laboratory in the Directorate of Agriculture. Later under the initiative of the Dewan of Mysore Sir M. Vishweshwaraiah, the Mysore Agriculture Residential School was established in 1913 at Hebbal which offered Licentiate in Agriculture and later offered a diploma programme in agriculture during 1920. The School was upgraded to Agriculture Collegein 1946 which offered four year degree programs in Agriculture. The Government of Mysore headed by Sri. S. Nijalingappa, the then Chief Minister, established the University of Agricultural Sciences on the pattern of Land Grant College system of USA and the University of Agricultural Sciences Act No. 22 was passed in Legislative Assembly in 1963. Dr. Zakir Hussain, the Vice President of India inaugurated the University on 21st August 1964.

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2008 - 200912010 - 20151
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Author: SATISH KUMAR, K. × End date: 2015 × Type: Thesis × Thesis Type: M.Sc ×
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    ThesisItemOpen Access
    CLONING AND EXPRESSION OF HEPATITIS B SURFACE ANTIGEN (HBsAg) GENE IN E. coli, Pichia AND Coleus forskohlii
    (UNIVERSITY OF AGRICULTURAL SCIENCES GKVK, BENGALURU, 2015-07-19) SATISH KUMAR, K.; RAMANJINI GOWDA, P. H
    Hepatitis B virus (HBV) infection is a worldwide health problem, which can lead to severe liver disease mainly hepatocellular carcinoma and cirrhosis. Hepatitis B surface antigen (HBsAg) is a glycoprotein found on the surface of the virus. The existing third generation vaccine is a recombinant Hepatitis B surface antigen (rHBsAg) produced in Yeast system (Saccharomyces cerevisiae or Pichia pastoris). Available vaccine is costlier, hence low cost production of vaccine is necessary to reduce the incidence rate especially in developing countries. Enhanced expression levels in yeast system or alternative search for host system for low cost production of vaccine is necessary to meet the demanding need of low cost vaccine. With these constrains in view, the present investigation lays emphasis on expression of codon optimized HBsAg gene (coHBsAg) in E. coli, Pichia pastoris (Intracellularly and Secretory) and Coleus forskohlii. HBsAg gene was codon optimized and sub-cloned into expression vector E. coli (pET28a), Pichia (pPICZA and pPICZ A) and transferred into E. coli BL21 and Pichia pastoris X-33 respectively. The positive clones were confirmed by DNA sequencing and restriction enzyme analysis. E. coli clone (pET28a_HOP-C1) showed yield of 1.56 mg/L, Pichia non-secretory clone (pPICZA_HOP-C10) recorded yield of 8.67 mg/L, Pichia Secretory clone (pPICZ A_HOP-C5) yielded 11.85 mg/L. Recombinant protein isolated from different host system was characterized by SDS-PAGE and Western blotting. Fused recombinant protein isolated from E. coli and Pichia non-secretory showed size of 25.4 kDa, Pichia secretory and plant system showed band of size 34.7 and 24.0 kDa respectively. Purified protein isolated from different host systems were used for immunization studies in albino mice. Results showed that E. coli expressed HBsAg showed highest titer value of 2.43 and plant produced HBsAg recorded second highest with value of 1.70 on par with positive control (1.22).
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    ThesisItemOpen Access
    CLONING AND CHARACTERIZATION OF rubisco PROMOTER FROM SUNN HEMP (Crotalaria juncea L.) FOR ITS ACTIVITY IN EXPRESSING FMDV GENES IN HOMOLOGOUS SYSTEM
    (University of Agricultural Sciences GKVK, Bangalore, 2008-11-26) SATISH KUMAR, K.; SURYANARAYANA, V .V. S.
    Genetically engineered microorganisms are important sources of industrial and medicinal proteins. Over the past decade Plant host system has been investigating as potential host system for expressing proteins of therapeutic and diagnostic use. However concerns regarding the stability and environmental safety need to be addressed for taking up the Plant system as potent host system. Chloroplast engineering however is expected to resolve some of the issue. Since, the FMD is a major concern in the world over and attempts have been directed towards development of alternative vaccines for conventional virus based immunoprofilactics, so present investigation used FMD immunogen for genetic engineering of chloroplast DNA and constructing transfer vector for transformation of Sunnhemp a fodder crop through Biolistic method. Using pBS-KS+ vector as a back bone rbcL promoter (1Kb) along with a 5’ end of gene with MCS (375bp) was amplified from genomic and chloroplast DNA and cloned in BamHI and HindIII, and BamH1 and SacI respectively, region that are highly conserved and flank the gene of interest and helps in homologous recombination. To the constructed transfer vector prbcL Trans, FMDV immunogenic gene P1-2A (2.3Kb) was cloned in the MCS BamHI and NotI. The plasmid was transferred into DH5α (E. coli) cells and the cultures were harvested at different time periods and the proteins from the lysates were analyzed for FMDV specific gene P1-2A. SDS-PAGE analysis showed an addition band of 90KDa indicating there was an expression of FMDV gene under cloned promoter. The specificity of the expression protein was confirmed by western blot. The constructed vector with immunogenic P1-2A gene (prbcL P12A) was used for transforming Sunnhemp calli standardized which induced a good callus from leaf explants in MS basal media with 2.0mg/L BAP + 0.2mg/L 2,4D through Biolistic method. The callus subject to bombardment with plasmid coated tungsten particles was co-cultivated for 15 days. The callus survived was further subjected to PCR for screening of the putative transgenic callus. The PCR was done using VP1 specific primer which amplify 700bp VP1-2A region. Two out of seven calli showed positive amplification resulting in a putative transgenic sunn hemp calli.