“However, the DNA-dependent RNA polymerase proved elusive until 1960, when it was independently identified in bacteria by Hurwitz and Stevens and in plants by the Bonner group” (p. 8325). There were also many experiments after these in which the scientists focused mostly on the mechanisms of the RNA polymerization (Kireeva, 2013). Kireeva also stated in her article that a group called the Chamberlin group focused on the pauses by E. coli RNA polymerase the group also focused on the…
Brains were sectioned at 16 µm in a cryostat, mounted on Super-Frost Plus slides with embedding medium and stored at -80℃. Signals of ISH were detected by digoxigenin (DIG) labeled single strand cRNA probes. The DIG-labeled cRNA probes were transcribed in vitro from cDNAs of rat GR (~0.4 kb) and rat SERT (~0.7 kb) in pGEM-3Zf vectors with T7 RNA polymerase and DIG RNA labeling mix (Roche, Branford, CT, USA). The amount of the transcript was estimated by comparison to DIG-labeled control RNA…
eukaryotes is that genetic information is formed in protein and is converted into organization, function and cellular structure. Eukaryotes also have a replication of machinery that is found in bacteria but are more complex and larger. The phase of replication has more factors for helicase and primase complexes, it then loads onto a sliding clamp unit where polymerase is there as…
joint pain, liver inflammation, and decreased hearing capabilities among other difficulties. Those who survive can no longer transmit the disease. Negative sense, single stranded RNA (ssRNA) constitutes the virus’s genome. It is considered to be a negative sense because the RNA cannot be directly translated into functional proteins. It must first be transcribed into mRNA, which is complementary to the ssRNA, before it can be translated. The ssRNA is contained in virions that are cylindrical or…
transcriptional machinery of RNA Polymerase I in the oocyte Experimental Approach: Aim1: To address the roles of the Hira complex in rRNA transcription and embryonic development The Hira histone chaperone complex is an evolutionarily conserved complex, which is composed of Hira, Ubn1 and Cabin1, and cooperates with the histone chaperone Asf1a to mediate H3.3-specific binding and chromatin deposition12,13. I previously demonstrated the critical roles of maternal Hira /H3.3 on paternal…
a. Introduction Lujo Hemorrhagic Fever is caused by a bi-segmented negative RNA virus that is one of several viruses known to cause viral hemorrhagic fever. The Lujo virus is part of the Arenaviridae family, which consist of several viruses categorized into two groups. The Lujo virus is categorized in the “Old World” group, which signifies that the virus in question was found in the Eastern Hemisphere, specifically the continent of Africa. The majority of the arenaviruses categorized in the…
In both prokaryotes and eukaryotes, it is a complex multistep process that’s regulated by a variety of enzymes and proteins. Prokaryotic and eukaryotic DNA replication are both bi-directional processes, the DNA polymerases work from the 5’ to the 3’ end, they both involve leading and lagging strands and primers are required2. The main differences result from the difference in complexity between both types of organisms. While prokaryotic DNA is circular, eukaryotes have linear chromosomes with…
Nucleoli are comprised of DNA, RNA, and proteins and disappear during prophase of cell division only to reassemble during telophase [5]. In addition, nucleoli consist of two separate parts: the granular component and the fibrillar component. The fibrillar component contains a large quantity of RNA polymerase I, which transcribes ribosomal RNA, and the granular component is where the later stages of ribosome production take place [5]. Besides its function to produce ribosomes, some scientists…
75 ml of buffer PE to the samples and spun it for another minute then disposed of the waste. The columns were added to the tubes and spun for a minute in the centrifuge to dry then planed in new clean tubes. Without touching the wafers with the pipet tip, 50µl of elution buffer EB was added to the columns then left for a minute and centrifuged for a minute. The liquid collected at the bottom of the tube is the purified DNA and it was placed on ice. In vitro Transcription: To recreate the…
identifies a specific sequence on the DNA and makes a “nick” or cut at that location. The nick is created by the protein and allows the DNA to unwind and continue the process. Once the DNA opens forming an open complex from there a lagging strand will develop along the 3’ template strand due to the polarity and orientation of the strands. A leading strand will emerge and DNA polymerase enlists nucleotides to pair with the exposed DNA template while RNA primer provides a 5’ end for nucleotides to…