When Daughter Chromosomes Go Through Recombination The

The purpose of this experiment was to determine the reversion rate and recombination rate of two mutant T4 bacteriophages, rII 29 and rII 31. Through recombination rate, the map unit between the two mutants was calculated.

Titer of the Bacterial virus, the Measurement of the Recombination and the Reversion Rate, and the Gene Map Distance (Phage Recombination)

134). They are loops of DNA that are separate from the chromosomal DNA and can self-replicate in a cell, found mostly in bacteria (Brown, 2011; Addgene, 2015). Lederberg and William Hayes discovered that plasmids were being transferred from one cell to another, not the chromosomal DNA (Brown, 2011, p. 135). This discovery lead to plasmids being an essential tool for scientists. Scientists can engineer plasmids to have specific genes to introduce into new cells (Brown, 2011, p. 134). On a plasmid loop there will be an origin of replication (ORI) and a multiple cloning site (MCS) where the gene of interest is inserted (Bio-Rad, 2015). This region has specific restriction enzyme recognition sites, which are cut by the enzymes to open up the DNA where the new gene will be inserted (Jove Science Education Database, 2015). Most plasmids will also contain an antibiotic resistance gene allowing cell survival in environments containing antibiotics (Jove Science Education Database, 2015).

In this case, the F is a dihybrid with AB on one chromosome and ab on the other

This process of P-element mobilization is the basis of how we directed recombination in male flies, and therefore the basis of generating deletions in the DMAP1 gene. We made several crosses that allowed P-element mobilization to occur in male flies by crossing P-element strains (non-autonomous) to a transposase source, producing heterozygous flies containing a P-element and a transposase source, and therefore allowing us to induce and detect male recombination events in progeny. The specific recombination event of interest is when the P-element is mobilized towards the right onto a homologue, which may have induced a deletion in

One of the most imperative functions in maintaining the development of evolution is the frequency of genetic transformation: the injection of foreign DNA into another organism’s DNA. This term is defined by the actions of a vector, but more specifically by the actions of plasmids and phages. However, in this experiment we are primarily focused on the effect of the pGLO plasmid transformation of GFP on the E. coli bacteria by introducing a second chromosome or a plethora of cloned plasmids. (Bassiri 2011)

After recognizing the restriction sites, the restriction enzymes cut off the DNA strands at these points, leaving sticky ends on the original DNA fragment. The restriction enzymes can also travel to other cells to cut off DNA sequences, the sequences that can match the bases of the original sequence. The sticky end will find other sticky end to form a new recombined DNA sequence, obeying the law of base-pairing.

· Describe how a piece of foreign DNA can become a part of a plasmid that ends up altering the final phenotype of cells

The transposon in this experiment is contains kanR in between the inverted repeats on either end, which will be transposed from the plasmid pVJT128 to the chromosome of the recipient bacteria.

On September 12, 1016, Belmont University graciously allowed Dr. Katherine Friedman from Vanderbilt University to come and talk to a crowd of students about the tendencies of how deoxyribose double stranded breaks can during cell replication and the elements required to hopefully repair this ordeal. She began the session by discussing what chromosomes are composed of and how they are produced, accompanied by visual and statistical representations. Moving on, she touched on how double strand breaks are a huge threat to a cell's, an organisms, stability. Correspondingly, she described what can cause these breaks; chemical factors, as well as inner cell disruptions during replication that are sometimes hard to remedy. However, she also stated that this breaks can occur on purpose, mostly in the immune system in efforts to make antibodies.

After the replication fork has been established the strands of DNA are ready for the next stage. On each strand is a

Plasmids are small double stranded circular non chromosomal DNA molecules containing their own origin of replication. Hence, they are capable of replication independent of the chromosomal DNA in bacteria. Plasmids present in one or more copies per cell, can carry extra chromosomal DNA from one cell to another cell and serve as tools to clone and manipulate genes. Plasmids used exclusively for this purpose are known as vectors. The genes of interest can be inserted into these vector plasmids creating a recombinant plasmid. Recombinant plasmids can play a significant role in gene therapy, DNA vaccination, and drug delivery [Rapley, 2000].

Interestingly, studies from our laboratory and others have demonstrated a stereochemical dependence for dSp processing by enzymes in ODNs. The first instance of a dSp diastereomer difference was reported from our laboratory when Klenow Fragment (exo-) was allowed to insert dATP opposite the isomers for which dSp1-containing ODNs were found to be the more favorable substrate for insertion and bypass.32 This study was extended to in vivo experiments by Neeley, et al. to include pol II and pol IV, also showing more favorable bypass kinetics for dSp1.34 As further evidence for in vivo stereochemical dependency in dSp processing, Henderson, et al. utilized the restriction endonuclease and postlabeling assay (REAP) to determine the mutation profiles for dSp1 and dSp2 in Escherichia coli.28 In the sequence context 5’-d(GSpA)-3’, they found a strong stereochemical dependence in the relative amounts of dG→dT and dG→dC transversion mutations for dSp1 and dSp2. In subsequent experiments, Delaney, et al. showed a somewhat different dependence of mutation profiles on the dSp diastereomers in the sequence context 5’-(TSpG)-3’ using the REAP and competitive replication of adduct bypass assays (CRAB).29,30 An additional observation from our laboratory quantified the digestion kinetics of dSp isomers in the dinucleotides 5’-d(Np[Sp])-3’ (N = A, T, G or C) by nuclease P1 that showed dSp1 was the kinetically preferred substrate in all sequence combinations.33 Lastly, in collaboration

After a BP or LR recombination reaction, this cassette is replaced by the gene of interest to generate the entry clone and expression clone, respectively.

Plasmid DNA with Restriction Digest: The purpose of restriction digest of plasmid DNA is to understand how each DNA plasmids was cut with the given restriction enzymes and perform gel electrophoresis to observe the samples. Nine restriction digests were created, containing three digests for each of the three plasmid DNAs identifying as recombinant, non-recombinant, and unknown. Out of the nine digests, six are actual digests and three are undigested controls. A master mix is created to add to each of the nine samples with its following stock ingredients: 10 ul of 2X Reaction Buffer, 1 ul of Nco1, X ul of sterile water (Single digest), 10 ul of 2X Reaction Buffer, 10 ul plasmid DNA, 1 ul Nco1, 1 ul of Not1, and X ul of sterile water (Double