I need the answers for (e) (f) (g) (h) (i) Question 2 (50 points) The template s
ID: 215903 • Letter: I
Question
I need the answers for (e) (f) (g) (h) (i)
Question 2 (50 points) The template strand of a human DNA and a plasmid are given below. left to right. Both DNAs are to be digested by the four base pair cuter in the table. The strands are read from Human DNA: 3'.. ACCATGGCAGGTACCGAACCTCAACTTTCCTC-5 Restriction Enzyme Name Sequence and type of cut 2 Bi GATC-3 -CTAG5 5- CGA-3 Ci Di Ei 4 3 CAT G-5 5-AG CT-3 Number each DNA from left to right with the first base numbered "1" and determine the a. cutting sites of the restriction enzymes on both the human and bacteria DNA. b. Which restriction enzyme(s) can be used in cloning the human DNA into the bacteria? Explain? c. What will be the total number of fragments in both the human and bacteria DNA d. What kind of ends will be produced after the cut, a cohesive or blunt end? e. How many nucleotides are in each fragment? (label each fragment according to number of nucleotides contained in the fragment) Describe what will happen if the restriction enzyme in (b) is used to cut the DNAs and all the fragments from both DNAs are mixed with the uncut DNAs of both species and agarose gel electrophoresis is conducted on the mixed fragments. Show the bands that will form on the agarose gel. f. g. Assuming the fragments of the human DNA is to be cloned into the bacteria, describe h. If a bacteria cell is to be transformed with the recombinant DNA, which kind of library i. Determine if the bacteria cells will express any protein from the human DNA. If yes, how this could be achieved will be formed? what kind of amino acid chain will be produced (don't worry about promoters, enhancers and transcription factors in the recombinant plasmid, just consider only the fragments cloned into the plasmid and assume that any genes can be transcribed from the fragment).
Explanation / Answer
e. Number of nucleotides in
i. Bacterial DNA - 40 nucleotides
ii. Human DNA- 43 nucleotides.
f. Restriction nuclease 'b' is 5' GATC-3'
3' CTAG-5'.
There is no sequnce in both bacterial and Human DNA with this sequence of 'GATC' .
g. The fragment of DNA is inserted into ampicillin resistant gene of E.coli and the transformant consists of human DNA in its plasmid and will express itself in the bacterial cell.
h. If the bacterial cell is transformed with recombinant DNA, we get a genomic library. A genomic library is a collection of the total genomic DNA from an organism. Construction of geomic library involves creating many recombinant DNA molecules.
i. Human DNA fragments inserted in bacterial cell will express in the bacterial cell and produce a protein called recombinant protein.