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
First isolated from a rhesus monkey in Uganda, the Zika virus (ZIK V) did not pose a serious global threat until recent outbreaks in 2007 on Yap Island in Micronesia and the current outbreak which began in 2013 in the French Polynesia. And, although the virus is transmitted via infected mosquitoes, studies indicate that the disease potentially can be transmitted sexually (Musso, D., Roche, C., Robin, E., Nhan, T., Teissier, A., and
In this investigation pUC19 plasmids were used as the vector due to its small size of 2686bp, high uptake efficiency by the host and fast replication time. Important features of this plasmid include the origin of replication and multiple cloning sites (MCS). The origin of replication allows the plasmid to replicate inside the host bacterium. The MCS is located within the lacZ gene and contains unique sites for the Xbal & EcoRI restirction enzymes to cut and produce sticky ends for the CIH-1 gene to bind to. Furthermore, the pUC19 plasmid also contains an ampiccilin resistance gene so only transforemed E.coli are able to remain viable when spread on the agar plates that also has the addition of ampiccilin. The lacZ gene encodes the β-galactosidase enzyme which aids in indentifying the recombinant E.coli from the non recombinant cells (Coventry University 2016).
Zika virus was discovered by scientists in the Zika rainforest in a remote part of Uganda in 1947, in the serum of an infected rheseus monkey. With five thousand two hundred thirty- four known cases of the infectious virus documented in the United States. There is currently no vaccination for the flu like virus. The chief concern of the healthcare administration is to continue to take preventive measures to prevent contracting the virus. Within the same flavivirus family
A multidisciplinary team from the University of Texas Medical Branch at Galveston is the first in the world to engineer a clone of Zika virus strain. The findings were published in a study on Monday. This development can expedite many aspects of Zika research including
As of now, there is no official vaccine for Zika. Instead, a patient that has the contagion must undergo certain treatments. Fortunately, the remedies for the virus are simple and cheap. If infection occurs, it is vital to get an abundance of rest and liquids; medication is also advised to assist in alleviating pain. The treatments for Zika are straightforward because the disease itself is rather mild. In fact, in most reported cases of the virus in the United States, the illness only lasted approximately two to seven days prior to the first set of symptoms (“World Health Organization”). It is evident that the fear associated with the Zika Virus is slightly exaggerated. Currently, there is only one recorded fatality in the United States that is directly related to Zika (“The Washington Post”). This is mainly because first world countries’ medical programs are advanced enough to effectively treat the virus. The standard of living in these countries are also higher compared to third world countries. That is why Zika has claimed a significant amount of lives worldwide; the poor conditions in these countries make it difficult for infected individuals to get proper
February 1, 2016 the World Health announced a PHEIC (Public Health Emergency of International Concern) that declared the Zika virus was a global health emergency. Brazil being the most affected country in the world right now has cases where pregnant women with the virus have given birth to children with smaller heads causing their brains to not develop correctly.The Zika virus is a flavivirus (Karimi) that is related to the common dengue virus and West Nile disease. A flavivirus is a group of RNA (a ribonucleic acid), that are mostly carried by mosquitoes, flies, fleas, etc. The virus carried causes severe human diseases, that are most commonly related to the Dengue and West Nile disease. The disease
Neural progenitor cells are permissive of Zika virus (ZIKV) infection in humans, and the ZIKV’s ability to infect such cells is a major global concern, because no other virus from the Flaviviridae family can infect brain cells. AXL was identified as the virus’s mediator for entry and disruption of radial glial cells and endothelial cells of the brain. To analyze the virus- host interaction, specifically the reprograming of the host proteins directed by AXL after Zika entry, the study of the effect of ZIKV infection on the epigenome in mouse radial glial cells is to be performed. To visualize the epigenetic changes caused by the virus the mapping of the epigenome of both infected and mock RGC cells is done. Whole ZIKV-infected E17.5 fetal brains
Zika virus epidemic in the Americas is linked to increased prevalence in microcephaly in infants infected with the Zika virus (ZIKV) in utero. The purpose of the study was to develop a vaccine to for women of childbearing age and their sexual partners to prevent infection and Viremia due to ZIKV. The authors created a DNA vaccine that can produce pre-membrane and envelope proteins that offer immune protection in mice and primates. This study showed that DNA vaccine is an effective way to protect against ZIKV.
The zEDIII is the domain of the E protein which binds receptors and contains epitopes of neutralizing antibodies against ZIKV. In this multi step experiment conducted by Yang (and others) they found characteristics of zEDIII such as its potent immunogenicity, its refolding and purification factors, and its lack of ADE(antibody dependent enhancement) which make it a promising vaccine hopeful if they can enhance/confirm its safety. In this experiment they concluded that complete protection against the ZIKV comes from a purified inactivated ZIKV, a plasmid DNA, and an E protein (prM-E). Scientists have the foundation, but it is going to take much more research to ensure no harm would come from this potential vaccination. This experiment demonstrated for the very first time that the immunization of recombinant zEDIII protein (different genetic material) provided a strong antigen specific response to ZIKV neutralization which is correlated to protection from the virus demonstrated in mice. They are making huge steps towards a Zika vaccination. Vaccinations need to very precise before they can be available to the public because they will be injecting the virus to produce that immunity against the virus which can be very
Given the increase in viral replication and number of infected cells within the placenta in the presence of cross-reactive DENV-elicited antibodies (Fig. 2E-F), we sought to identify the ZIKV-infected cells within the placenta in the presence or absence of enhancing antibodies. Several previous studies, in both mice and humans, have identified placental macrophages (Hofbauer cells) and trophoblasts as potential in vivo targets for Zika virus (ZIKV).(19-25) To evaluate ZIKV infection in macrophages, we co-stained for ZIKV NS3 and the macrophage marker MAC-2. Immunofluorescent co-staining of ZIKV-infected placentas from mice receiving CTRL plasma revealed approximately 3% of macrophages to be ZIKV-infected (Fig. 3A, C). Mice receiving DENV-immune plasma showed a significant increase in the total number of ZIKV-infected macrophages,
As Zika migrated across the globe, its symptoms seemed to progress with its travel. As the virus traveled it mutated and evolved to include a vast array of different symptoms and accelerated its ability to affect more populations. In today’s society, the symptoms of Zika are a lot more severe and more prevalent than ever in the past. Scientists link this rise in severity and prevalence of symptoms to an increased number of transmission routes and symptoms. Thirty years ago, we did not see the neurological symptoms that we see today. Zika was thought of as just a mild
Throughout the Caribbean, Central, and South America there has been health crisis that’s has been affecting the youngest generation of these developing countries. Thousands of babies have been born with a birth defect known as microencephaly. This birth defect can cause horrible problems such as seizures, mental retardation, developmental delays, short stature and difficulties with coordination and balance. (Centers for Disease Control 2017) This birth defect often leads to having to provide life-term medical care and it was determined to be caused by the mother acquiring a mosquito-borne infection with the Zika virus. So, what is this Zika virus and how can
As stated in the beginning of this paper, CHIKV has similarities in signs and symptoms with Dengue Virus. Scientists were trying to formulate a more sensitive and specific approach to appropriately diagnose CHIKV. Currently, scientists are developing ways to better differentiate Zika, Dengue and CHIKV as they bear similarities in signs and symptoms. Despite having different genus species, they are similar in envelope protein folding and membrane fusion mechanisms. Furthermore, scientists studied proteomic sequencing provided by the National Center for Biotechnology Information (NCBI) protein database and researched on polymorphisms, potential protein fragments and possible antigenicity of peptides, and finally, calculated amino acid
This article discusses the causes and characteristics of ZIKA viral disease. The main focus of the article is on the identification of ZIKV inhibitors and demonstration of the efficiency of the viral polymerase inhibitor
Zika virus ZIKV is a flavivirus that has several modes of transmission across humans. Classically, the mosquito is the vector of transmission and yet, more recent literature shows that the outbreak of ZIKV viruses were sexually transmitted from infected individuals to uninfected individuals. Concerned for the general health of the population, Lim et. all attempt to understand the overall enhancement of ZIKV infection by preexisting antibodies against other flaviviruses, more specifically, against dengue (DENV) and West Nile virus (WNV). The importance of this research allows the scientific community to ensure that vaccines created against a specific virus will not cause more harm when infected by another flavivirus, considering the