Why is it important to include the start and stop DNA sequences for the jellyfish Glo protein?
We did make sure to include a start and stop dna sequences for the jellyfish so that the Glo gene would be transcribed and expressed and so we can see that we have successfully transformed the cells into which we place the engineered plasmid.
Why do you need to use the same restriction enzyme to cut the plasmid DNA and the gene of interest?
Restriction enzymes cut at specific sequences so the same restriction enzyme must be used because it will produce fragments with the same complementary sticky ends, making it possible for bonds to form between them. Their sticky ends match, and so they can be ligated together.
Why can DNA fragments cut with the same restriction enzyme bind together?
The principle is simply that, if two different DNA molecules are cut with the same restriction enzyme, both will produce fragments with the same complementary sticky ends, making it possible for DNA chimeras to form.
Why is it important to find an enzyme that would cut the plasmid at one site?
Why was it important to discard any enzymes that cut the plasmid at the replication site? (If the plasmid were cut at the replication site, it would not be able to reproduce and transfer genetic information to its host bacterial cell.) In this activity, you incorporated an insulin gene into the plasmid.
What enzyme does the tape represent?
The tape represents the enzymeligase that affixes the new gene into the plasmid. This new plasmid with recombinant DNA is ready to be reinserted into a bacterium so it can make growth hormone.
Why do you only want the plasmid to be cut once?
1. Why was it important to find an enzyme that would cut the plasmid at only one site? What could happen if the plasmid were cut at more than one site? You simply want to open the circular DNA so that the human DNA can be inserted into the circle.
Why do the sticky ends pair and stick together?
Very often they cut the two DNA strands four base pairs from each other, creating a four-base 5′ overhang in one molecule and a complementary 5′ overhang in the other. These ends are called cohesive since they are easily joined back together by a ligase.
Why are blunt ends less useful?
Compared to sticky-end ligations, blunt-end ligations are less efficient, in fact, 10 – 100 times less efficient. This is because, unlike sticky end cloning, there is no hydrogen bonding between the complementary nucleotide overhangs to stabilize the formation of the vector/insert structure.
What enzyme glues the new DNA strand together?
DNA Ligase
What is DNA complementary strand?
Complementary DNA (cDNA) is a DNA copy of a messenger RNA (mRNA) molecule produced by reverse transcriptase, a DNA polymerase that can use either DNA or RNA as a template. From: Encyclopedia of Genetics, 2001.
What 2 scientists built the first DNA model?
The 3-dimensional double helix structure of DNA, correctly elucidated by James Watson and Francis Crick.
What enzyme is responsible for unzipping DNA?
Helicase Key
What is the main function of Primase?
Primase is an enzyme that synthesizes short RNA sequences called primers. These primers serve as a starting point for DNA synthesis. Since primase produces RNA molecules, the enzyme is a type of RNA polymerase.
What enzyme removes the RNA primers?
DNA polymerase I
What enzyme is responsible for splitting the two strands quizlet?
The first step in DNA replication is to separate or unzip the two strands of the double helix. The enzyme in charge of this is called a helicase (because it unwinds the helix).
What are the bonds that are broken to break it into two strands?
In general, DNA is replicated by uncoiling of the helix, strand separation by breaking of the hydrogen bonds between the complementary strands, and synthesis of two new strands by complementary base pairing. Replication begins at a specific site in the DNA called the origin of replication.
Which enzyme is responsible for facilitating?
Answer: The DNA polymerase is responsible for facilitating the hydrogen bonds between nucleotides in a new DNA molecule.
What kind of bonds are broken as the DNA unzips?
Explanation: Helicases are enzymes involved in unzipping of the double stranded DNA molecule at beginning of DNA replication. They do so by binding at DNA sequences called origins on DNA molecule then they break the hydrogen bonds between complementary base pairs causing the two strands of DNA molecule to unzip.
What happens after DNA unzips?
DNA replication occurs through the help of several enzymes. These enzymes “unzip” DNA molecules by breaking the hydrogen bonds that hold the two strands together. Each strand then serves as a template for a new complementary strand to be created. Complementary bases attach to one another (A-T and C-G).
What is unzipping DNA called?
The first step in DNA replication is to separate or unzip the two strands of the double helix. The enzyme in charge of this is called a helicase (because it unwinds the helix). Once the strands are separated, an enzyme called DNA polymerase copies each strand using the base-pairing rule.