Materials And Methods Of Protein Sequences Of Lassa Virus

* Flaviviruses: share a common size (40-60nm), symmetry (enveloped, icosahedral nucleocapsid), nucleic acid (positive-sense, single stranded RNA approximately 10,000-11,000 bases), and appearance in the electron microscope. Therefore, images of West Nile virus are representative for this group of viruses.

The filovirus Ebola consists of 4 families: Marburg, Ebola Zaire, Ebola Sudan, and Ebola Reston. It is a rather simple virus in structure; each contain a single strand RNA strand and 7 different proteins, 3 which are only slightly understood and 4 that are completely unknown. The virus harms the immune system like the HIV virus, but Ebola causes an explosive attack. The virus is associated with the measles and mumps family, pneumonia viruses, parainfluenza viruses which include colds, and respiratory system related viruses. Unlike common viruses which are ball-shaped, Ebola’s shape represents that similar to a shepherd’s crook, a long snake-like thread that looks like a worm. Because the Ebola virus has a distinct structure, it has been classified as a “thread virus” in its own family of viruses, the filoviruses.

Nipah virus, Arena Virus and Francisella tularensis are bioterrorism agents. They work in various ways to harm the host. Francisella tularenis is considered a Tier 1 bioterrorism agent and Nipah virus is an overlap select agent that affects humans and agriculture both. They have been harmful in the past. Though, Nipah virus is a newer virus than the other two. This review will focus on their emergence, pathogenicity and symptoms of the diseases they cause.

In this experiment, the primary antibodies against DAT were generated from the species Rattus norvegicus (the lab rat). The secondary antibodies (Chicken Anti-RatDAT) synthesized against Rat Anti-DAT, were generated from Gallus gallus domesticus (chickens). The Chicken Anti-RatDAT antibodies were synthesized by injecting certain purified rat antibodies into a chicken. As a result, a generation of the monoclonal antibodies Rat Anti-DAT, was essential to detect the DAT segment of the protein. Likewise, the polyclonal antibody Chicken Anti-RatDAT, linkage to the Fc portion of Rat Anti-DAT. Secondary antibodies are known to assist in the sorting, detection, and/or purification of target antigens by attaching to a primary antibody, that directly attaches to the target antigen.

First, since we know the viral RNA sequence and its targeting protein, we can investigate it in bioinformatics database, and can acquire some clues or hints about the target proteins. From the bioinformatics database, we might be able to find its structure, similarity with other proteins, functions, and binding domains. In other words, we can get some partial or complete amino sequences of the targeting proteins or information about likeness.

One of the most complex and unexplored disease causing agents are viruses. They are known to be able to infect a wide array of organisms, from plants and animals, to bacteria and fungi. Essentially, anything that is living is capable of being infected with a virus. Once the host's cell is infected, the viruses' goal is to produce more viruses which will infect neighboring cells to continue the cycle.

The predicted antigenic regions were chimerically expressed in a bacterial system. Briefly, two sets of PCR primers were designed according to gD gene sequence available in GenBank (accession number: NC_001847) to amplify gene fragments encoding aa 20-160 (nucleotide 118953～119375, Δ gD1) and aa 257-344 (nucleotide 119664～119927, Δ gD2), respectively. The primers sequences for amplification of Δ gD1(P1 and P2) and Δ gD2 (P3 and P4) were listed in Table. 1.

Hendra and Nipah viruses from genus Henipavirus are paramyxovirus that can cause diseases that passed between animals and humans. These viruses are responsible for causing diseases that occurs at irregular intervals at some places. Like other paramyxovirus, henipavirus also have the ability to escape the host immune response due to interferon protein; this is due to P gene. The P-gene is responsible for encoding four proteins – C, P, V, & W protein. Currently no vaccines or therapeutic remedies are available which can prevent henipavirus infection. Ribavirin which is used to treat various viral hemorrhagic fevers, in vitro studies of ribavirin against both hendra and nipah virus shows effective inhibition of hendra and nipah virus. The henipavirus have two envelope proteins F & G glycoproteins; the G glycoprotein is an attachment glycoprotein while F is the fusion glycoprotein. The recent tactic to prevent henipavirus infection focused on direct medication by soluble forms of F & G viral glycoproteins to induce a protective immune response.

According to Points of View Reference Center, “3000 Miles to a Cure’s mission is to end brain cancer by raising funds for research, extending an open invitation to unite in hope and action for the cure(smith-strickland).”This quotation means people are funding brain cancer and running for research for scientist. In order to find a cure for brain cancer, people need to look for cures,fund research,run for awareness,so scientist and doctors can make a breakthrough in history.

Lastly, they also stimulate apoptotic genes in order to cause cell death. Memory T cells are able to clone themselves and remain in circulation to prevent reinfection. So, if pathogens such as L. pneumophila reappear, cytotoxic T cells will immediately be reactivated to fight off infection. Helper T cells stimulate T and B cells, and suppressor T cells inhibit the function of both T and B cells. T cells have receptors on their surface, also known as TCRs. They are capable of recognizing antigens bound to glycoproteins in plasma membranes. There are two classes of MHC proteins. Class I is found in the membranes of all nucleated cells. Class II can be found in the membranes of antigen-presenting cells. T-cell glycoproteins allow the epitope to bind to the TCR. CD8 are associated with cytotoxic T lymphocytes, and CD4 are associated with helper T lymphocytes. CD8 recognizes MHC I proteins, and CD4 identify MHC II proteins. MHC I proteins gather small peptides in cells and bring them to the surface. Abnormal proteins or peptides activate T cells, which destroy the cell. MHC II proteins recognize antigenic fragments, activating T cells to fight off foreign cells and

This homotetramer protein with 97 amino acids in each monomer comprises a short region in N-terminal ecto domain, a transmembrane domain and a long protein molecule which have ion channel activity and regulate vesicular and cytoplasmic pH within the virus-infected cells [32, 33]. M2 ecto domain which is known as M2e, include 24 amino acids from N-terminal which remained highly conserved. Moreover, another region of the N-terminal which includes 2 to 9 amino acids that is famous as SLLTEVET epitope is conserved among all influenza A subtypes. Considering the existence of such conserved epitopes in M2, this region seems to be a reasonable choice for designing the universal vaccine [34]. Nevertheless, the point is that M2e conserved region is not large enough to trigger immune response alone. This problem has been solved by fusing different types of molecular adjuvants like HBc, ASP-1, CTA1-DD and bacterial flagellin [35-38].

Chikungunya is a small spherical enveloped virion that measures up 60 to 70nm in diameter. It contains a single strand positive-sense RNA genome (Solignat, Maxime 2009). The virus has a lipid bilayer containing heterodimeric protein spikes composed of two major surface envelope glycoproteins E1 and E2 (Understanding the alpha virus). The genome extends approximately 12 kb long, and is organized in a 5’ cap -untranslated region (UTR), non-structural proteins (nsP1-4), structural proteins (C-E3-E2-6 K-E1), and a 3’ polyA tail end( Microbes and Infection). The structural proteins of the virus are created by the translation of a messenger RNA (mRNA). The 5’ end cap of the genome has a 7- methyl guanosine cap, whereas the 3’ end is polyadenylated

The immense diversity of T-cell repertoire, the universe of pathogenic epitopes, and the polymorphism, codominance of HLA-alleles makes the identification of antigenic T-cell repertoire of any pathogen daunting. Several advances in genomics, proteomics and computational techniques have resulted in the development of novel approaches for identifying the HLA class I-restricted antigenic repertoire of CTLs in the past decade. For instance, biochemical methods such as mass spectrometry (MS) can perform unbiased identification of the HLA-ligandome of any cell type (Kowalewski et al., 2015; Shao et al., 2017). However MS-identification while being sensitive, does not identify immunogenic epitopes that result in T-cell activation, and generally requires large amounts of sample. Fluorescent HLA-multimers in combination with multiparameter flow cytometry can be used identify immune reactive CTLs, and have recently been combined with genomics to identify immunogenic

An in-silico study used immunoinformatic approach to find the heights antigenic MERS-COV protein, found that N protein pose the highest antigenic score and vaccine against the N protien can elicit both neutralizing antibody and cellular immunity against MERS-COV infection while vaccine against the S protein of MERS-COV can elicit only a neutralizing antibody, the study mapped T-cell epitopes, B-cell epitopes and CTLs epitopes to N protein to find putative epitope vaccine candidates. B-cell epitope was excluded due it's lower

This essay based on the principle of real time PCR which uses CYBR green dye that combines to any double stand DNA. This process included two maim steps. The first step was designing of HSV-1 primer and /or HSV-2 primer (we have chosen only HSV-1). BioEdit software has been used to edit the nuclide sequences where necessary. After that the primer has been ordered. The second step was in the laboratory which included applying the HSV-1 primers to real time PCR protocol. The final results of this protocols aim to demonstrate the quality of HSV-1 primer that we have designed by interpretation three criteria. These are specificity, efficiency and