UNIVERSITY OF AGRICULTYRAL SCIENCES, DHARWAD

DEPARTMENT OF GENETICS AND PLANT BREEDING

Masters Seminar on

Plant Molecular Farming

NAME: Manjunathswamy N Hiremath DATE: 22.11.2013 ID No: PGS12AGR5846 TIME: 2:00 PM .......SYNOPSIS

Plant molecular farming (PMF) is a new branch of plant biotechnology, where plants are engineered to produce recombinant pharmaceutical and industrial proteins in large quantities. Various transformation strategies have been known to be used but transient expression remains best [3]. Vitamin A deficiency causes symptoms ranging from night blindness to leading to total blindness. Because no rice cultivars produce this provitamin in the endosperm, recombinant technology was used to introduce this pathway. Immature rice endosperm is capable of synthesizing the early intermediate geranylgeranyl diphosphate, with the use of 4 enzymes or through bacterial carotene de-saturase the -carotene can be synthesized. Three constructs were used where in one was for single transformation Pb19hpc & remaining for co-transformation pZPsc & pZLcyH, further HPLC analysis shown that co-transformed once were producing higher amount of carotene [6]. Human immunodeficiency virus type 1 (HIV-1) has infected more than 40 million people worldwide. One means of antibody production is the use of plants. HIV-1 Pr55Gag is a promising HIV-1 vaccine candidate: they compared the expression of this and a truncated Gag (p17/p24) and the p24 capsid subunit in Nicotiana spp. using transgenic plants and transient expression. Transient agro infiltration was best for expression of all of the recombinant proteins tested, and p24 and p17/p24 were expressed at much higher levels than Pr55Gag [1]. Oral administration of vaccines remains the only economically feasible approach to mass immunization of humans on a global scale. Therefore, the DNA fragment encoding hepatitis B virus surface antigen was introduced into plants to obtain transgenic lupin (Lupinus luteus L.) and lettuce (Lactuca sativa L.) expressing envelope surface protein. Mice that were fed the transgenic lupin tissue developed significant levels of hepatitis B virusspecific antibodies. Human volunteers, fed with transgenic lettuce plants expressing hepatitis B virus surface antigen, developed specific serum-IgG response to plant produced protein [2]. The E7 oncoprotein from Human Papilloma Virus (HPV) is an attractive candidate for anti-cancer vaccine development. In this study, they engineered HPV16 E7 coding sequence (wild type or mutagenized sequence, E7GGG) as fusions to -1,3-1,4-glucanase (LicKM) of Clostridium thermocellum and produced in Nicotiana benthamiana plants using a transient expression system. Target antigens were evaluated in mice for their potential as prophylactic and therapeutic vaccine candidates. Both fusion proteins induced E7-specific IgG and cytotoxic Tcell responses and protected mice against challenge with E7-expressing tumor cells [5]. An effort to produce an edible vaccine for rabies, the rabies glycoprotein gene is transformed into cucumis melo Var. NS-Abhijit, through Agrobacterium mediated transformation. The presence of the transgene was confirmed through PCR analysis. Western blots confirmed the specificity of this foreign protein. The extract of the fruits when inoculated in to mice produced Substantial levels of neutralizing antibodies [4]. Key Words: PMF Plant Molecular Farming , HIV Human immunodeficiency , HPV Human Papilloma Virus
References:

1. Ann Meyers., Ereck, C., Enid, S., Fiona, L. T., James, M., Alisson, L., Anna, L. W. and Edward, P. R., 2008, Expression of HIV-1 antigens in plants as potential subunit vaccines. BMC Biotechnology, 8: 115. 2. Kapusta, J., Modelska, A., Figlerowicz, M., Pniewski, M., Letellier, L. O., Yusibov, V., Koprowski, H. A., Plucienniczak and Legocki, A. B., 1999, A plant-derived edible vaccine against hepatitis B virus. FASEB, 13: 17961799. 3. Olawole, O., Obembe, J. O., Popoola, A., Sadhu, L. B. and Siva, V. R., 2011, Advances in plant molecular farming. Biotechnology Advances 29: 210222. 4. Ramanjini Gowda, P. H. and Madhusudhana, S. N., 2001, Production of rabies glycoprotein in musk melon (var.ns-abhijit) through agrobacterium mediated transformation and testing for its efficacy in experimental mice. Official J. of the Association for Prevention and Control of Rabies in India (Regd.), 2: 108-112. 5. Silvia, M., Rosella, F., Rossella, B., Antonio, M., Vadim, M., Vidadi, Y. and Aldo, V., 2007, Vaccine Anticancer activity of plant-produced HPV16 E7 vaccine. Vaccine, 25: 30183021. 6. Xudong, Y. P. B., Salim, A., Paola, L., Patrick, S., Ralf, W. and Ingo, P., 2000, Engineering the Provitamin A (b-Carotene) Biosynthetic Pathway into (Carotenoid-Free) Rice Endosperm. Science, 287: 303-305.