dna biology and technology

24.1
Hershey and chase experiment
● They want to determine if protein or dna (deoxyribonucleic acid) is the genetic material.
● They knew that viruses are okay because it has tiny particles which consists of inner nucleic core and an outer nucleic
core called a capsid.
–Experiment 1
•Viruses with DNA labeled with 32P were incubated with E.coli
–Mixed in a blender to remove virus particles attached to cells
–Centrifuged so bacteria formed a pellet
•Results- viral DNA was inside the bacteria
–Experiment 2
•Viral proteins in capsids were labeled with 35S and viruses were incubated with E.coli
–Mixed in blender and centrifuged
•Results- labeled proteins were washed off with the capsids and were not inside the bacteria
● Therefore, dna is the genetic material cause it stays inside.

Structure of dna
● Deoxyribonucleic acid
● By watson and crick in early 1950s.
● A double helix
● A polynucleotide and each nucleotide has,
1. Phosphoric acid (phosphate)
2. Pentose sugar (deoxyribose)
3. A nitrogen containing base
● Four bases
1. Purines with a single ring -Adenine (A) and Guanine (G)
2. Pyrimidines with a single ring - Cytosine © and thymine (T)
Complementary base pairing
● Dna forms a double helix. The strands of a helix are held together by hydrogen bond between the bases.
● A binds with t with two hydrogen bonds
● c binds with g with three hydrogen bonds or vice versa

● Dna strands are antiparallel because they run in opposite directions. (3’ to 5’) and (5’ to 3’)
Replication of dna
● Dna replication is the process of copying one dna helix into two identical helices
● It is semiconservative because each new double helix has one old conserved strand and new strand. Meaning ->
parental strands is present in each new double helix.
● Because each old strand produced a new strand through
complementary base pairing , there are now two
identical dna helices.
1. Before replication begins, the 2 strands of the parent
molecule are hydrogen-bonded together
2. Enzyme dna helicase unwinds and “unzips” the double-
stranded DNA
3. New DNA nucleotides fit into place along divided
strands by complementary base pairing
4. New nucleotides form bonds with the existing ones-
DNA polymerase
5. DNA ligase repairs any breaks in the sugar-phosphate
backbone
6. Two daughter DNA molecules have now formed that are
identical to the original

24.2

Structure of RNA
● Ribonucleic ACid
● Composed of nucleotides
1. Phosphate
2. Ribose (SUgar)
3. Nitrogen containing base
● Adenine and guanine
● Cytosine and uracil ( uracil replaces thymine)
● –3 major classes of RNA
•mRNA- carries genetic information from the DNA out to ribosomes
•rRNA- composes ribosomes, site of protein assembly
•tRNA- brings in amino acids to the ribosomes
-there is atleast 1 tRNA found in every 20 amino acids

Gene expression (production of protein ) requires two things : transcription and translation

Transcription
● RNA polymerase (transcription enzyme) attaches to a promoter, which is a segment of DNA nucleotides at the beginning of a
gene.
● The enzyme breaks the Hydrogen bonds between the nitrogenous bases and will read the 3’ to 5’ DNA template strand.
● RNA polymerase will add RNA nucleotides (A, U, C, G) to the DNA template strand by complementary base pairing. If DNA base is
adenine, it pairs with T and if DNA base is C, the complement is G.
● The mRNA then detaches and the Hydrogen bonds between the DNA base pairs reforms.
● When the gene has been read, an mRNA is formed and it runs in a 5’ to 3’ direction.

● Exons segments of mRNA that do code for amino acids; are joined together during mRNA processing. Only exons
result in protein product.
● Introns segments of mRNA that do not code for amino acids in the polypeptide; removed during mRNA processing.
● RNA polymerase transcription enzyme; breaks Hydrogen bonds between nitrogenous bases of gene; complements
with RNA nucleotides; check for correct complement and corrects if necessary; reforms Hydrogen bonds within a gene
(refer to transcription)

Genetic code
● Triplet code: 3 RNA nucleotides grouped together in an mRNA referred to as “codons”; codes for a specific amino acid that will be
placed in a specific polypeptide
● There are 64 mRNA codons . but there are only 61 triplets that correspond to an amino acid,the remaining three are
stop codons (uaa,uag,uga)
● Start codon” methionine “ (Aug) signals polypeptide initiation.

MULTIPLE CHOICE

% of a base in a DNA sample will equals to 100%; complementary bases will equal each other; for example, [A (35%) – T (35%) =
70%] + [C (15%) – G (15%) = 30%] = 100%

Hershey and Chase experiment 1952; used bacteriophages (viruses), bacteria, radioactive phosphorus and radioactive sulphur; proved
that DNA (not protein) is the genetic material; since protein coat remains outside the bacteria, DNA must carry the hereditary
infos

Franklin and Wilkins- double helix provided an x-ray diffraction photograph (to Watson and Crick) that determined the
structure of DNA is double helix

Ribose/Deoxyribose sugars Ribose-RNA, w/ O2; has one more oxygen atom // Deoxyribose-DNA; w/o O2; has one less oxygen atom.
Complementary base pairing DNA = A-T (double covalent bond), C-G (triple covalent bond); RNA = A-U, C-G

Steps of DNA replication unwind double helix by breaking H bonds by enzyme, DNA helicase (1), Exposed DNA strands form a
replication fork (2), [Leading Strand] DNA polymerase reads template strand in 3’ to 5’ (3). Enzyme adds DNA nucleotides (4) by
complementary base pairing. Initiated by RNA primer and added to origin of replication (5).

● mRNA codon sequence RNA nucleotides sequence; grouped in triplets; each codes for a specific a2 that will be placed in a
specific polypeptide
● DNA helicase breaks the H bonds between the bases/ unzips the double helix
● DNA polymerase pastes DNA nucleotides by complementary base pairing to the template strand
● Triplet code 3 RNA nucleotides grouped together in an mRNA referred to as “codons”; codes for a specific amino acid
that will be placed in a specific polypeptide
● Exons segments of mRNA that do code for amino acids; are joined together during mRNA processing
● Introns segments of mRNA that do not code for amino acids in the polypeptide; removed during mRNA processing
● Translation ribosome reads or translates the genetic message encoded in the mRNA codons in order to synthesis a
polypeptide; chain initiation, elongation, termination
● RNA polymerase transcription enzyme; breaks H bonds between nitrogenous bases of gene; complements with RNA
nucleotides; check for correct complement and corrects if necessary; reforms H bonds within a gene
● RNA promoter a segment of DNA nucleotides at the beginning of a gene; site where RNA polymerase attaches
● Transposons specific DNA sequences that have the ability to move within and between chromosomes; their movement
sometimes alters neighbouring genes by increasing or decreasing their expressions; one of the causes of mutations
● mRNA function carries genetic message for protein synthesis to a ribosome
● mRNA processing (posttranscriptional) Segments of mRNA that do not code for amino acids in the polypeptide
(introns) are removed and segments of mRNA that do code for amino acids (exons) are joined together. An altered
Guanine nucleotide (cap) and a poly-A tail are then added. The nuclear pore will determine the time it takes an mRNA
to enter the cytoplasm.
● PCR requires Taq polymerase, DNA nucleotides and DNA primers
● Taq polymerase DNA polymerase from a bacterium that lives in hot spring (Thermus aquaticus)
● Thermus aquaticus hot spring
● Heterochromatin inactive genes are present; darkly stained; wound around histone proteins
● Euchromatin active genes is present; loosely packed
● Transcription activators attaches to the enhancer, which is at a distance from the promoter; transcription factors
unite them to the RNA polymerase; allows RNA polymerase to attach to the promoter.
● Frameshift mutation- NOT is not a point mutation; insertion of RNA nucleotide or type and order of amino acids
change after deletion
● Requirements for rDNA formation plasmids (vectors), from E. Coli, transport a foreign gene into bacterial cells by
transformation. An enzyme/molecular scissors (restriction endonuclease) then cuts both the foreign and plasmid
DNA at specific DNA nucleotide sequence sites to create sticky ends (regions where foreign DNA join to plasmid DNA by
complementary base pairing). DNA ligase then bonds the sticky ends of foreign DNA to those of the plasmid.
● Restriction endonuclease – functions enzyme/molecular scissors the cut both foreign and plasmid DNA at specific DNA
nucleotide sequence sites to create sticky ends
● Operon – researchers F. Jacob, J. Monod, & A. Lwoff (discovered the control mechanism of gene transcription in E.
Coli; they studied the production of three enzymes used by bacteria to digest lactose)
 ● Protein activity (posttranslational) polypeptide takes a specific shape; activity of many proteins is short-lived;
proteins are degraded by proteasomes
●
● DNA-RNA-Protein
● DNA to mRNA
● DNA to DNA
● Transgenic animal use
● Operator function
● Regulator gene function
○
● Compare prokaryotic mRNA with eukaryotic mRNA (2)
● Discuss the procedure for inserting a foreign gene into a plant cell OR an animal cell (2)

Translation
● Initiation:
○ Small ribosomal subunit attaches to the mRNA start codon (AUG).
○ Methionine-tRNA (start codon) then, attaches with a complementary codon-anticodon match.
○ The large ribosomal subunit then attaches to the small ribosomal subunit to form a ribosome.
○ Met-tRNA is now in the P binding site.
○ A2-tRNA entered the “A” binding site with a complementary codon-anticodon match (H bonding occurs)

● Elongation:

o Ribosome will then continue to read mRNA codon until a stop codon enters the A site.

Termination Events when Stop Codon is in the A binding site (1)

● Translation will end with a stop codon (UAG, UAA or UGA) as no anticodon complements it.
● A releasing factor (protein) enters the A site and the ribosomal complex separates (polypeptides, tRNA, mRNA and
ribosomal subunits detach from each other).
Why do proteins take a specific shape during termination? (1)

● Proteins takes a specific tertiary or quaternary (contains two or more polypeptide) in order to be a functional protein
during termination.

Importance of an Anticodon to a tRNA (1)

● Anticodon in tRNA tells us what amino acid is attached to the CCA 3’
Nuclear Pore Function in Protein Synthesis (1)
● mRNA, tRNA and ribosomal subunits migrate through nuclear pores to enter the cytoplasm.
Relationship between Nucleolus and rRNA (1)
● Within the nucleolus, different rRNA molecules will combine with various proteins to form either the small or the
large subunit of a ribosome

Discuss DNA replication of the lagging strand given six terms (3)

● The DNA polymerase will read the lagging strand in reverse direction (3’ to 5’ direction).
● RNA primers will be added to specific locations called origin of replication along the lagging strand.
● DNA polymerase then adds DNA nucleotides, called the Okazaki fragments (A, T, G and C) by complementary base pairing
(A-T & C-G) between the RNA primers when given the “green flag”.
● DNA ligase will link the Okazaki fragments together to produce a continuous, complementary strand
Why was the one gene-one enzyme hypothesis changed? (1)

● It was discovered that some enzymes consisted of two or more polypeptides linked together and the synthesis of each
chain is controlled by a different gene. This changed one gene-one enzyme hypothesis to one gene-one polypeptide
hypothesis.
Gene mutation results in a codon change (2)

● When the first or 2nd base of a codon is changed, a DIFFERENT amino acid is usually added
● When the 3rd base of a codon is changed, the SAME a2 is usually added.
Discuss “system on” in the Lac Operon (3)

● Lactose is present and it binds to the repressor proteins and this prevents the lac repressor from attaching to the
operator gene (O).
 ● RNA polymerase then bonds to the promoter gene.
● (O) is signaled to turn on transcription in the structural genes
● mRNAs are then read by different ribosomes and the lactose-digesting enzymes are synthesized.
Explain mRNA processing (2)

● Segments of mRNA that do not code for amino acids in the polypeptide (introns) are removed and segments of mRNA
that do code for amino acids (exons) are joined together.
● An altered Guanine nucleotide (cap) and a poly-A tail are then added.
● The nuclear pore will determine the time it takes for an mRNA to enter the cytoplasm.
Differences between two-point mutation types (2)

● Point #1, #2 & #3 are also called substitution

● Point #1 & #3 is when the first or second base of a codon is changed and usually a different amino acid is added.
● Point #2 is when the third base of a codon is changed and the same amino acid is usually added.