Could you please explain the Q 4 and 5? Highlighted in pink marker. 4. The uniqu
ID: 85927 • Letter: C
Question
Could you please explain the Q 4 and 5? Highlighted in pink marker. 4. The unique properties of the restriction enzymes.. this means that 'EcoRI has AATTC sequence'?! 5. Plasmid should have an antibiotic resistant gene (selective marker)--what this mean? Also, how do we select the transformed bacteria with the plasmid inside them? Thank you so much!GLO and DNA elect ho hegad DNA fragments move in an electrical field. Large fragments move slowly. DNA is negatively charged. Review the questions discussed in extra credit handout. the Ara ow p xperiment and the questions that were discussed in the lab. Know operon that is a part of this plasmid. Plague Forming Assay method xpo 6-3). What is the Burst size? Recombinant paper DNA u You should know the terms: plasmid, restriction endonucleases, transformation 2 What is the goal of the recombinant DNA technology? and genetic engineering? What is a How does it apply to DNA sequences 4 You should know the unique properties of the restriction enzymes For gene cloning (recombinant pa plasmid that we performed in the lab), know the important points to consider when cloning a gene. Such as, Plasmid should have an antibiotic resistant gen The restriction enzyme should not cut withi the gene; otherwise gene would be inactivated. How do you select the transformed bacteria with the plasmid inside them? Hamburger iable Plate Count Know the Serial Dilution techniqu (Exp 6- How to calculate Dilution Factor and The in a sample.
Explanation / Answer
4. Since restriction enzymes cut DNA only at specific target sequences and not randomly, they are suitable for manipulation of DNA i.e. for use in recombinant DNA technology. The EcoRI restriction enzyme recognizes only G^AATTC sites. Hence the targeted DNA gets inserted into the plasmid when it is cut with the EcoRI restriction enzyme and the plasmid DNA is also cut with the same enzyme as this generates sticky ends that are complementary and ligated.
5. The plasmid that transforms a bacterial cell has, origin of replication, a polylinker i.e. multiple cloning site (MCS) which contains many (up to ~20) restriction sites and antibiotic resistance marker gene. The transformed cells when grown in the presence of the antibiotic are viable and able to grow and form colonies and are hence selected over the nontransformed that are not able to thrive in the presence of antibiotic.