Plantas transgênicas de tomate expressando o gene do antígeno PfCP‑2.9 de Plasmodium falciparum

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.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

Malaria Vaccine Candidate Diversity Offers Challenges and Opportunities for Effective Vaccine Development

Malaria is one of the most deadly diseases caused by protozoan parasites of genus Plasmodium. It affects 300-500 million people annually, of which more than a million lives are lost; among them majority under 5 years of age. By conventional wisdom, the immune mechanisms responsible for protection against malaria will require a multiple of 10-15 antigen targets for proper protection against various stages of malarial infection. Such large number of targets cannot be delivered to humans, by this method. Moreover, each antigen is reported to be highly polymorphic in nature and the malaria-affected populations live in economically poor part of the world. Development of anti-malarial vaccines is therefore, a very tough challenge from technical, delivery and affordability points of view. Technical challenges include identification of epitopes / antigens against appropriate targets, construction of DNA vector(s) that will express properly folded functional protein. Vaccine delivery challenges include developing an easy method to deliver multiple doses within a short period of time to infants and children of less than five years of age. Affordability challenges include development of cost-effective vaccines that can be stored at room temperature and be easily delivered. Although the complex life cycle of Plasmodium is challenging for anti-malarial vaccine development, it also offers a lot of antigen targets (opportunities) to combat malaria. Information on anti-malarial vaccine candidates, DNA constructs and cost-effective delivery mechanism will be discussed