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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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Author: SATISH KUMAR, K. × End date: 2015 × Start date: 2010 × 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).