Charlie Scheftic

BME 463
HW 5
6.12
a. D = ug = (um*S)/(KSX*X + S)
S = (D*KSX*X)/(um D)
b. Given the equation from a) and D(S0 S) = (ug*X)/(YMX/S)
S = (S0* YMX/S*D*KSX)/(um D + D*KSX*YMX/S)
c. S will double
8.2
Methionine can be overproduced by altering the regulatory mechanism through
transformation and transduction. It can also be overproduced by providing high
concentrations of amino acids to prevent the biosynthesis of amino acids by
feedback inhibition. Another way to overproduce methionine would be to use
mutants that are resistant to methionine so they would not experience feedback
inhibition and repression. The last way to overproduce methionine would be to
modify the pathway in a way that encourages production if one enzyme is shut
down.
8.7
The source of the DNA would likely be the lac Z gene which codes for Betagalactosidase (the color changing protein). Regulatory elements like promoters (lac
promoter) should be used in order to promote transcription and the protein making
process. These promoters can also interact with repressors to turn off the gene
when needed. The donor DNA will be inserted into the vector after the vector is cut
at the proper restriction site by a restriction enzyme. The vector is then injected into
E. coli through transformation which involved treating the E. coli with high
concentrations of calcium ions with temperature manipulation. You would then
screen the gene with the cells on a medium containing lactose and penicillin. Since
the cells contain a penicillin resistant gene, the cells that will be used for production
are alive and producing blue colored colonies.
8.9
a. The three methods are shotgun cloning, hybridization, and the intron splicing
method (cDNA). Shotgun cloning makes use of restriction enzymes to cut
DNA into fragments. Hybridization makes use of short fragments (probes)
that are complementary to the gene. Using introns, specific sequences can
but cut out and transfected into out genetic material like vectors.

b. The best method to use would be the intron splicing method (cDNA) so that
the processed mRNA can create complementary DNA strands through reverse
transcriptase and produce large amounts of human proteins using bacteria.