RESULTS AND DISCUSSION PCR and dsRNA production Target sequences of 1501, 1576, 1650, 1750, 538, and 716 bp specific for gyrase A, gyrase rpo B1, rpo B2 and GFP were successfully amplified with PCR (data not shown).. The in vitro transcription using T7 enzyme resulted in target specific dsRNA of 1501, 1576, 1650, 1750, 538, and 716 bp for each of gyrase A, gyrase B, rpo B1, rpo B2, L11, and GFP genes, respectively (Fig.1). In vitro growth inhibition assay B. bovis growth (Fig. 2) from an initial parasitemia of 1% was significantly (ANOVA) inhibited at 10 µg/ml and 50 µg/ml concentrations of gyrase A, gyrase B, rpo B1 and B2, and L11 (Fig. 2). Treatment with a mixture formed of 10 µg/ml gyrase A and 10 µg/ml gyrase B significantly inhibited the growth (ANOVA, P < 0.01). The mixture formed of 10 µg/ml rpo B1 and 10 µg/ml rpo B2 resulted in significant inhibition of the growth (ANOVA, P < 0.01) (Fig. 2). The single treatment with 10 µg/ml or 50 µg/ml of each of the dsRNA of gyrase A, gyrase B, rpo B1, rpo B2, and L11 (Tukey-Kramer HSD) did not show significant difference. There was no significant difference between the single treatment with 10 µg/ml or 50 µg/ml of each dsRNA and the mixture formed of 10 µg/ml gyrase A and 10 µg/ml gyrase B dsRNAs and the mixture formed of 10 µg/ml rpo B1 and 10 µg/ml rpo B2 dsRNAs
Now that the Plasmid ‘psiRNA’ is transfected into the somatic cell, scientists don’t need to inject dsRNA into the cell each time to stop the production of BLG protein, the plasmid automatically produces siRNA into the cytoplasm
Throughout the cells there are much type of Eukaryotic cells which make many types of cells and then they proceeds to the RNA, There is no complete data of this RNA present and the characteristic are very poorly unstated. The genetic information is directly represented by the RNA and it focuses on its synthesis, translation and modification it helps to understand the genome functions also. These observation are taken up together to form and define the functions and description related to genes (Bell, 2004). This observation tells about the range of expression and localization. As the technology is been improving day by day for the RNA profiling and the type of isolation made by the cells , the number of RNA has grown and
To develop nursing career in an environment that offers tremendous potential for professional growth and achievement; exploring opportunities offered with impactful challenges utilizing my experience, skills and passion.
Since shRNA does not interact with the Drosha/DGCR8 microprocessor complex, this RNA must be designed with care in order to achieve correct downstream processing and gene suppression.1 Proper dosage of shRNAs is another factor to consider, as it has been found that they may saturate the endogenous pathway and disrupt natural miRNAs.4
Each 1.5 ml microcentrifude tube containing 50 µg myceliums, 50 µl of Lysozyme, 450 µl of Lysis solution and 3 µl of RNase A was kept for hour at 37 0C on water bath, vortex several times in duration. Then, addition 250 µl of 2% SDS, gently invert before addtion 250 µl of mixing Phenol- Chloroform- Isoamylclcohol ratio is 25:24:1, vortex and centrifuge at 12,000 rpm in 10 minutes then transfer supernatant on up layer to a new 1.5 ml microcentrifuge and repeat that step again before the next step to completely remove unexpected components. Subsequently, 0.1X of 3M Sodium acetate pH 4.8 was supplemented,
revealed the existence of abundant amount of microsporidia in the bloodstream of H.axyridis under light microscope and again confirmed using electron microscope. Once microsporidia enters the bloodstream, it undergoes RNA transcription inside the cell. But in H.axyridis it is in inactive state due to absence of gene transcription factor and doesn’t cause any harm in them perhaps it has established immunity towards them. This forms a shield for H.axyridis eggs as well as larvae but when ingested by native ladybird species i.e. Coccinella beetles leads to death. Amplification of microsporidia shows that its RNA matches closely with Nosema thomsoni. This is present in all H.axyridis by vertical
B. bigemina, B. caballi, B. bovis, and T. equi were gained from cultures with parasitemia of 5%, which was diluted with suitable uninfected red blood cells to a starting parasitemia of 1% for the assays. The growth inhibition assay was done in 96-well plates containing 20 μl of packed red blood cell inoculum and 200 μL of an appropriate culture medium containing 5, 10, 25, 50, and 100 μM of ebselen. The concentrations used were based on a initial study. Positive control cultures received 5, 10, 50, 100, 1000 or 2000 nM of diminazene aceturate (Aboulaila et al., 2010a). For negative experimental control, cultures without the drug and cultures containing DMSO 0.01 % for Ebselen and DDW 0.02 % for diminazene aceturate were arranged. The experiments were carried out in triplicate wells per drug concentration for each parasite species and in three isolated trials. Cultures were incubated at 37 °C in an atmosphere of 5 %
Agarose GE: Figure 1 shows the results of amplifying five types Arabidopsis thaliana fwa gene. successful PCR was a result of nice bands showing in figure 1. Wild type undigested (900bp), mutant undigested(860bp), and mutant digested (850bp) were all showing band in the same location. On the other hand, Wild type digested did not show any bands meaning amplification did not occur because McrBc digests fwa enzyme that is being methylated. Gel electrophoresis was used to determine the size of the base pairs from the logarithmic equation. Digested and undigested mutant genes showed hazy regions meaning they have less amount of amplification compared to wild type undigested which shows a significant amplification. No band was shown for
The small amount of colonies were put into a labeled PCR tube which contained: 10 µl GoTaq Master Mix, 1 µl F primer for the SSU, 1 µl R primer for the SSU, and 7µl of H20. The tubes went through a various thermal cycler program. 10min at 95 celsius, 30 seconds at 95 celsius, 30 seconds at 55 celsius, 2 minutes at 72 celsius, and 10 minutes at 73 celsius. The tubes were placed in a refrigerator until gel electrophoresis.
spiral is inside every nucleus of body cells. The number of body cells is approximately 60 trillion.
DNA isolation is a process of DNA purification through physical and chemical means. DNA was protected within the nuclear membrane of B. rapa, which was surrounded by a cell membrane and cell wall. Mechanical and chemical lysis of the cell was necessary to break open the cells and solubilize the membranes to isolate the DNA. Mechanical lysis, which consisted of grinding the leaves with a pestle, broke down the cell wall. Chemical lysis, through the use of lysis solution, broke down the lipid-base membranes. The lysate was placed inside the Zymo-spin IV spin filter to remove chunks of debris from the solubilized cell components. A binding buffer was added to the collection tube that held DNA,
Whereas the reaction products are not specific the SYBR green dye added to the reaction mixture to quantitatively indicate the amount of dsDNA in the reaction mixture by providing amplification plot and melt curve as well as the melt temperature plot that used in may offers useful information about the sensitivity of the DNA template in the sample (Giglio, Monis and Saint, 2003).
Due to the similarities between Plasmodium and Babesia and Theileria parasites luteolin might have inhibitory effect on these blood parasites. The aim of the present study was to evaluate microscopically the inhibitory effects of luteolin on the in vitro growth of three Babesia species and T. equi and on the in vivo growth of B. microti. Furthermore, to study the effects of luteolin on the transcription of B. bovis DNA gyrase subunits A and B by Reverse transcription-PCR
The Polymerase Chain Reaction is a method that uses the capability of DNA polymerase to synthesize to new DNA strands which are matching to the template strand. A primer needs to be added to the first nucleotide due to the fact that DNA polymerase only can add a nucleotide only onto a 3 '-OH group that already exists. Because of this condition, we are able to define a chosen region of template sequence which we can then generate millions to billions of copies. This technique was developed by Kary Mullis in 1983 and is a very common indispensable technique which has a variety of uses such as DNA cloning for sequencing, genetic fingerprints, detection and diagnosis of infectious disease (often cancer) etc...
Genetic modification, otherwise referred to as recombinant DNA (rDNA) technology or gene splicing, has proven to be more precise, predictable and a better-understood method for the manipulation of genetic material than previously attained through conventional plant breeding. Agricultural applications of the technology have involved the insertion of genes of desirable agronomic traits into a variety of crop plants, and from a variety of biological sources. Examples include soybeans modified with gene sequence from a streptomyces species encoding enzymes that confer herbicide tolerance, and corn plants modified to express the insecticidal protein of an indigenous soil microorganism, Bacillus thuringiensis. A growing body of evidence suggest that technology and may be used to make enhancements to not only the agronomic properties but the food, nutritional, industrial and medicinal attributes of genetically modified crops.