DNA ligase

three major types of DNA ligases are reported: DNA ligase I (hligI), DNA ligase III (hligIII) and DNA ligase IV (hligIV). hligI plays an important role in DNA replication by joining Okazaki fragments on the lagging strand of DNA. Apart from this it also plays important roles in DNA damage repair pathways, single strand breaks are repaired by nucleotide excision repair (NER) and base excision repair (BER) pathways. Results of preclinical studies support that Human DNA ligases are an attractive target

mRNA of the PY motif for ENaC(ref). This results in either a truncated protein or elimination of some amino acids from the PY motif, which interfere with the binding ability of Nedd4 ligase and inhibit Nedd4 ligase binding, respectively. Overall, this either decreases or eliminates the possibility of Nedd4-2 ligases’ ability to tag and degrade the ENaC. With the inability to control ENaC surface expression at the kidney, its reabsorption of sodium will be markedly increased due to

DNA cloning is the process of creating a multitude of copies of isolated DNA fragments; DNA cloning can be carried out via in vitro or in vivo methods. One can clone a specific DNA sequence or entire gene fragments. There are a multitude of procedures to carry out DNA cloning, but the major steps are the same for all types. To begin the process, one must isolate a DNA fragment from the chromosomal DNA. This is done by using a restriction enzyme. One could also use gel electrophoresis and polymerase

ampicillin in the agar solution was working. Reactions 1-3 were set up to locate the optimum ratio of bacteriophage insert DNA to create the most

achieve accuracy of DNA replication after the unwinding of DNA, the DNA must be synthesized. DNA synthesis, in the case, the two strands are disconnected and then turned into single-stranded DNA. This is called replication fork. The replication fork acts as a model for DNA synthesis. However, the site of impaired or mutated DNA can cause a lower rate pf success for DNA synthesis. Impaired DNA can also lead to unprocessed and unligated okazaki fragments. Okazaki frgaments are small DNA fragments that are

An important reaction that needed to be performed before introducing foreign DNA into E. coli is the ligation reaction. DNA ligation is a process that joins DNA fragments together by creating a phosphodiester bond between the 3’ OH of the nucleotide and the 5’ phosphate of another. An essential enzyme that ligates a DNA fragment and a vector together is the T4 DNA ligase, which originates from T4 bacteriophage. This enzyme will ligate cohesive ends produced by restriction enzymes. T4 will also ligate

Recombinant DNA is taking a section of DNA and placing it with another strand of DNA. The combination of two different organisms allows for a new strand of DNA. There are three ways in which recombinant DNA could be made. These are transformation, phage introduction, and Non-Bacterial Transformation. (David L. Nelson, 2012) In transformation, a piece of DNA is inserted into a vector. Then it is cut with an enzyme and the bonded DNA insert is placed in a vector that contains DNA ligase. The insert

a linear ladder was used to measure DNA segments that are in a nonlinear form. For ligation 1, there were supposed to see three bands present around 6.5kb, 5.5kb, and between 0.5kb and 1kb. However, there were no bands present. This could be due to multiple reasons, but the most credible source of error is blind/hover pipetting. Which most likely caused the pET-41a(+) vector and the egfp insert to not enter the solution at all. Thus, resulting in no form of DNA appearing in the ligation image. There

together to repair the DNA strand by joining together. There are two main types of DNA ligase, the first is found only in prokaryotic cells. The second is found in eukaryotic cells . Furthermore, mammals have four subtypes of ligases that vary in their function; DNA ligase III, this contains a DNA repair protein, called XRCC1, that helps to repair the DNA strand that occurs during nucleotide repair. Eukaryotic DNA ligases are larger than the prokaryotic cells. Therefore DNA ligase has a very important

The overall goal of the DNA lab was to create recombinant plasmids which we would use to alter the DNA of bacteria to make them ampicillin resistant. The first test of our success was the gel electrolysis. The gel photo allowed us to see if we had successfully ligated the recombinant plasmids. For the uncut samples, K- and A-, we expected to see two or three bands, since there are several forms of plasmids: supercoiled, nicked or open-circle, and the multimer. In K- there were two bands, and in A-