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Prvac59 Research Paper

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A particular stain of the Zika Virus, PRVABC59, has recently become an epidemic in South America and the Pacific¹, and in the last month has spread to Florida. PRVABC59 is a Flavivirus, ssRNA(+), is spread by Aedes Aegypti and Aedes Albopictus mosquitoes and is capable of spreading from mother to unborn child during pregnancy. Although the Zika virus normally doesn’t cause severe symptoms, further complications arise if a fetus is infected, as stated by the World Health Organization “There is scientific consensus that Zika virus is a cause of microcephaly and Guillain-Barré syndrome”. Before more research can be done to develop a vaccine or treatment, a specific primary antibody must be created as a tool for further research about the function of the ten proteins in the Zika genome. This project worked on nine of these proteins in ten parts. Two structural proteins, capsid and pre-membrane protein, and the seven non-structural proteins, with NS5 being split into two pieces due to its size and toxicity. A previous method of protein production and purification2 was followed to create these antibodies. This method of protein production and purification should lead to a primary antibody specific to recombinant proteins of the PRVABC59 strain of Zika virus, which is a necessary tool for further research into the PRVABC59 strain. …show more content…

Plasmid map of pRSETB. Primers were designed to amplify via PCR to cDNA. The PCR product was digested with XhoI and EcoRI enzymes and ligated into the pRSETB plasmid. The pRSETB plasmid contains a T7 promoter region, is ampicillin resistant, inducible with Isopropyl β-D-1-thiogalactopyranoside IPTG, a molecular mimic of a lactose metabolite that triggers transcription of the lac operon, and has XhoI and EcoRI cut sites. The pRSETB plasmid is transformed into dH5α E. coli and plated on carbenicillin plates. Colonies are selected and grown on a carbenicillin plate while PCR is used to check that the plasmid that was up-taken was not

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