Transgenic tomato plants expressing the antigen gene PfCP-2.9 of Plasmodium falciparum

Abstract

-The objective of this work was to obtain transgenic tomato plants expressing the PfCP-2.9 protein (a chimera of the antigens MSP1 and AMA1 of Plasmodium falciparum). Cotyledons of seven-day-old tomatoes, cultivar Summers, were transformed via Agrobacterium tumefaciens. Transgenic expression in the T0 plants was verified in the DNA extracted from fruits. PCR analysis was used to test the presence of the gene of interest in the T1 generation. Reverse transcriptase PCR provided evidence of gene expression at the RNA level, and Western blot analysis confirmed the presence of the protein of interest in the T1 plants. This is the first report of successful transformation with the expression of a malaria antigen (PfCP-2.9) in transgenic tomato plants from the T0 and T1 generations.Index terms: gene expression, genetic transformation, malaria, plant-made vaccines. Plantas transgênicas de tomate expressando o gene do antígeno PfCP-2.9 de Plasmodium falciparumResumo -O objetivo deste trabalho foi obter plantas transgênicas de tomate que expressem a proteína PfCP-2.9 (uma quimera dos antígenos MSP1 e AMA1 de Plasmodium falciparum). Cotilédones de tomate, cultivar Summers, com sete dias de idade, foram transformados via Agrobacterium tumefaciens. A expressão transgênica nas plantas T0 foi verificada no DNA extraído dos frutos. A análise por PCR foi utilizada para testar a presença do gene de interesse na geração T1. A evidência da expressão do gene no RNA foi constatada por meio da PCR de transcriptase reversa, e a análise "Western blot" confirmou a presença da proteína de interesse nas plantas T1. Este é o primeiro relato de transformação bem sucedida com a expressão de um antígeno da malária (PfCP-2,9) em plantas transgênicas de tomate da geração T0 e T1.Termos para indexação: expressão gênica, transformação genética, malária, vacina a partir de plantas.

Tomato (Solanum lycopersicum L.) in the service of biotechnology

“…Notwithstanding, the available malaria vaccines produced by conventional methods are still unaffordable for most people due to their high price. Thus, Kantor et al (2013) initiated the production of antigen gene PfCP-2.9 of Plasmodium falciparum in tomato fruits. This research is the first report of a successful transformation with the expression of a malaria antigen (PfCP-2.9) in transgenic tomato plants of the T 0 and T 1 generations.…”

“…This research is the first report of a successful transformation with the expression of a malaria antigen (PfCP-2.9) in transgenic tomato plants of the T 0 and T 1 generations. Kantor et al (2013) found that transgenic tomatoes produced 35 mg per gram of fresh weight of leaves of malaria antigen protein. The TSP (Total Soluble Protein) extracted from the tomato leaves was similar to that obtained by other researchers (Biswas et al 2012).…”

Expression of Colorectal Cancer Antigenic Protein Fused to IgM Fc in Chinese Cabbage (Brassica rapa)

“…Expression of the EpCAM-IgM Fc protein in leaf tissues of transgenic plants was analyzed using Western blotting (Figure 4). In the IgM Fc detection immunoblot, all randomly selected transgenic lines (2,6,7,9,11) showed an approximately 75 kDa protein band similar to that of the positive control (Figure 4A black arrowhead). In the EpCAM detection immunoblot, all lines (2,6,7,9,11) of transgenic plants showed the expected bands for EpCAM-IgM Fc (Figure 4A) with an EpCAM-IgM Fc-sized protein band (Figure 4B white arrowhead).…”

“…Expression of the EpCAM-IgM Fc protein in leaf tissues of transgenic plants was analyzed using Western blotting (Figure 4). In the IgM Fc detection immunoblot, all randomly selected transgenic lines (2,6,7,9,11) showed an approximately 75 kDa protein band similar to that of the positive control (Figure 4A…”

“…Plant expression systems, such as those in tomatoes [1,2], carrots [3], bananas [4], spinach [5], lettuce [6], and tobacco [7,8] have been established to produce valuable recombinant proteins, including therapeutic enzymes, vaccines, and antibodies. Among horticultural crops, Chinese cabbage has been recently recognized as a potential candidate to produce such valuable recombinant proteins, because it has a reasonable total soluble protein capacity relative to plant biomass [9].…”