In Biotechnology, gene cloning is a very important technique. A vector is normally required to perform this process. The vector commonly used to transform a bacterial host cell is the plasmid. (i) State the THREE (3) important regions of the plasmid. Elaborate your answer.
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- List the advantages and disadvantages of using plasmids as cloning vectors. What advantages do BACs and YACs provide over plasmids as cloning vectors?In cloning human DNA, why is it necessary to insert the DNA into a vector such as a bacterial plasmid?If I clone a complete eukaryotic gene, including the eukaryotic promoter region, ligate it into a plasmid, and transform it into E. coli, will I be able use the transformed E. coli to make the corresponding protein? Explain why, or why not? If you decide to do this, what would your cloning strategy be?
- What is a plasmid? What are the minimum 3 components that a recombinant plasmid used for genetic transformation should have?What characteristics of plasmids makes them good cloning vectors? Discuss.You are attempting to clone a 3 kb gene from the bacterial sp Microbacterium foliarum into the EcoRI site of the 6.0 kb plasmid shown. If you restrict the plasmid with EcoR1, how many bands will you obtain on agarose gel electrophoresis? Compare it with a plasmid in which no gene has been inserted. Draw a representative agarose gel showing the bands, and the direction of current flow as well point out the positive and negative electrodes
- Which vectors (plasmid, phage λ, cosmid, bacterial artificial chromosome) can be used to clone a continuous fragment of DNA with the following lengths? a. 4 kb b. 20 kb c. 35 kb d. 100 kbPLease explain What characteristics of plasmids makes them good cloning vectors? DiscussYou have two samples you have to sequence: one is a cloned plasmid that you want to verify the sequence of, and another is the cDNA library of the transcriptome of a cell. Which method would you use for each sample and why?
- What is the most logical sequence of steps for splicing foreign DNA into a plasmid and inserting the plasmid into bacterium?I) Transform bacteria with recombinant DNA molecule.II) Cut the plasmid DNA using restriction enzymes.III) Extract plasmid DNA from bacterial cells.IV) Hydrogen-bond the plasmid DNA to non-plasmid DNA fragments.V) Use ligase to seal plasmid DNA to non-plasmid DNA A. III, II, IV, V, I B. I, II, IV, III, V C. III, IV, V, I, II D. II, III, V, IV, IBased on the picture, why plasmids are used as a vector for DNA Recombination? What other vectors can be used?A molecular geneticist hopes to find a gene in human liver cells that codes for an important blood-clotting protein. He knows that the nucleotide sequence of a small part of the gene is CTCGACTCACA. Briefly explain how to obtain the desired gene. Briefly describe how to clone the desired gene into a cloning plasmid.