Biochemistry Study Material with Solved Assignments and Essays - Desklib

Running head: BIOCHEMISTRY 1
Biochemistry
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BIOCHEMISTRY 2
PART A
1. Ribosomes require Aminoacyl-transfer RNA to read and interpret the genetic code.
2. Consider the following, in which two DNA fragments are treated with T4 DNA ligase: -
the first is a plasmid digested with restriction enzymes BamHI (sticky ends) and HpaI
(blunt ends) and not treated with phosphatase; and- the second is a BamHI DNA fragment
treated with phosphatase. Identify the possible outcome(s) of the ligation reaction:
Answer: a circular plasmid containing the DNA fragment
3. An intron is a short mutation in a non-coding region.
4. Which of these fermenter types does really exist? Shaken fluid-bed fermenter.
5. In genomic studies, gene identification allows us to understand proteome
composition.
6. Identify actual membrane transporter proteins on the list below. Antiporters, Symporters
along with ATP-coupled active transporters.
7. Alpha-complementation is a common method to verify the successful insertion of cloned
DNA fragments into a plasmid.
PART B
1. What are the following tools used for in recombinant DNA technology:
1. a) Hind III (50 WORDS)
It is a type II site-specific deoxyribonuclease restriction enzyme separated from
Haemophilus influenza. The tool is used to cleave the DNA palindromic sequence AAGCTT in
the cofactor Mg 2+ presence through hydrolysis (Tayi, Maku, Patel & Sonti, 2016). Hind III is
used for genetic engineering and molecular biology.
1. b) T4 DNA Ligase (50 WORDS)
It is a ligation enzyme utilized to join fragments of DNA through catalyzing the
formation of phosphodiester bonds amidst 3’hydroxyl termini and juxtaposed 5’phosphate in

BIOCHEMISTRY 3
double-stranded DNA by use of ATP as a coenzyme (Matsuzaki, Su, Walsh, Kennedy & Mei,
2018). Moreover, it seals nicks for the DNA substrates.
2. c) Calf intestine phosphatase? (50 WORDS)
Calf Intestine Phosphatase is utilized to catalyze the hydrolysis of 5’phosphate groups
from RNA, DNA and both ribo and deoxyribonucleoside triphosphates (Toro ET AL., 2018).
The removal of 5’phosphate is effective in avoiding self-ligation of the cleaved DNA vectors.
Furthermore, it dephosphorylates the DNA before kinase labeling reactions.
2. Answer the following with regard to DNA technology:
a) How and why you can separate and visualize DNA fragments by size? (50 WORDS)
DNA fragments are separated by size by a current run through a gel with molecules.
Depending on DNA size, the molecules move via the gel in opposed directions enabling them to
be segregated from each other. To visualize, the gel is discolored with a DNA binding dye and
put under UV light for the DNA fragments to glow.
b) Explain the principle behind the separation technique. (50 WORDS)
The principle behind the technique is to analyze DNA fragments produced by restriction
enzymes. It uses electromotive force to transfer molecules via a porous gel and separates them on
the basis of shape, charge, and size. However, the separation basis is grounded upon how the gel
and sample are prepared.
3. In recombinant DNA technology
a) What is a screenable marker gene used for? (50 WORDS)
A marker gene is utilized in molecular biology in ascertaining if a piece of DNA has been
effectively inserted into the host organism. A screenable marker gene is used to make the cells
carrying the gene be viewed differently (Bashir, Kim, Stahl & Cho, 2016).
b) Name two screenable marker genes. (50 WORDS)

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The green fluorescent protein (GFP) makes the cells glow green under UV light. Here, an
exclusive microscope is needed to view each cell (Sachsenhauser & Bardwell, 2018). On the
other hand, a GUS assay screenable marker gene detects a single cell by discoloring it blue
without using any complex apparatus.
4. With regard to PCR technology:
a) Outline the steps and main components of a PCR reaction; and (50 WORDS)
Steps in Polymerase Chain Reaction involve denaturing, annealing along with extending.
On the other hand, the basic elements of Polymerase Chain Reaction entail DNA template,
primer, DNA polymerase, thermal cycler, microfuge tube, Tris-HCl, MgCl2, Gelatin or bovine
serum, distilled water and deoxynucleotide triphosphates (Sochivko, Fedorov, Alekseev,
Kurochkin & Dubina, 2017).
b) Provide two examples of PCR applications. (50 WORDS)
PCR is used in forensic science for the recognition of lawbreakers along with the
gathering of organic crime scene proof like soil, semen, and blood. Moreover, PCR is applied in
clinical diagnosis to research into the diagnosis and help in treating various diseases. (Kim et al.,
2016).
5. Consider the fermentation process:
a) Name the main metabolites produced by lactic acid bacteria (LAB)(50 WORDS)
Lactic acid bacteria generate organic acids like propionic, acetic and lactic acids
generated as end products. Moreover, it produces antimicrobial metabolites like H2 O2 generated
during aerobic growth, bacteriocins which are ribosomally synthesized antimicrobial
compounds, ethanol produced through the heterofermentative pathway along with diacetyl
generated from excess citrate-derived pyruvate (Soro-Yao, Brou, Amani, Thonart & Djè, 2014).
b) Name five food products produced using LAB. (50 WORDS)