Fredrich Miescher
1869
First isolated or discovered DNA
Called it “nuclein” or nucleic acids
Fredrick Griffith
1928
Discovered “transforming factor” in bacteria
Harmless bacteria were transformed into harmful bacteria when introduced to dead, harmful bacteria
Oswald Avery
1944
Repeated Griffiths experiments
Discovered that the transforming factor in bacteria was DNA
Erwin Chargaff
1950
Determined
DNA differs between species
Pairing of bases in DNA
A = T and G = C
This is known as “Chargaff’s Rule”
Hershey & Chase
1952
Worked with bacteriophage viruses to prove viruses to prove DNA is the genetic material
Experiment Summary:
Radioactive sulfur stained viral protein coat
Radioactive phosphorus stained viral DNA
When virus infected bacteria, only radioactive phosphorus was found in the bacteria
Means DNA is the genetic material
Rosalind Franklin
1952
Used X-ray diffraction to produce “Photo 51”
Discovered DNA was double helix
Also worked with scientist Maurice Wilkins, who was the head of lab
Watson & Crick
1953
Also worked with scientist Maurice Wilkins, who was the head of the lab
Published work won Nobel Prize
Prokaryotic cells have…
1 circular DNA molecule
Eukaryotic cells have…
Strands of DNA in the nucleus
These strands become chromosomes
Chromosomes are made of…
Pieces of DNA coiled around proteins
These proteins are called histones
DNA meaning
Deoxyribonucleic Acid
DNA
Stores genetic information
Made of nucleotide
Shaped like double helix
Looks like twisted ladder
Sides are made of the sugar phosphate backbone
Alternating sugars & phosphates
Held together by covalent bonds
(Think of the backbone like the sides of the ladder)
The middle of the DNA molecule is made up of the nitrogenous bases
Like the rungs/steps of the ladder
Bases held together by hydrogen bonds
Nucleotide Parts
Sugar (deoxyribose)
Phosphate
1 of 4 nitrogenous bases
Adenine, Thymine, Guanine, Cytosine
2 sides of DNA molecule
Sides known as anti-parallel
Strands run from 5’ → 3’ direction
5’ always starts with a phosphate
Complementary strands run in opposite directions
2 types of nitrogenous bases
Purines and Pyrimaidines
Purine
Have 1 ring
Adenine and Guanine
Chargaff’s Rule
A = T and C = G
Because of this base pairing rule, DNA is said to have complimentary strands
About DNA
Chromosomes are made of DNA which contain sequences called genes
Genes code for various proteins
These proteins determine your traits
DNA is the code that provides instructions to tell your cells how to make proteins
The sequence of nitrogenous bases is the code that makes the particular protein for a given trait
Pyrimidine
Have 2 rings
Include Cytosine & Thymine
Uracil in RNA only
Always pairs with a pyrimidine
DNA Replication
The process by which DNA makes a copy of itself and in turn its chromosomes
Amount of Nucleotides in Human Body Cells
~6 billion pairs
How Many Hours to Copy DNA
6 hours
DNA Replication produces…
2 identical strands of DNA
The Base Pairing Rule Allows Each Strand…
To serve as a template
What Do The 2 Strands of DNA Contain
Complimentary info
Watson & Crick suggested that…
The DNA double helix separates and each new strand was copied
The Proposed 3 Models of DNA Replication
Conservative Model, Dispersive Model, Semi-Conservative Model
Conservative Model
1 original helix and 1 brand new helix produced after replication
Dispersive Model
Sections of original DNA mixed together with brand new DNA
Semi - Conservative
1 strand of original DNA and 1 strand of new DNA
Steps of Replication
The DNA unzips
Begins at specific sites called origins of replication
Copying moves out in both directions creating replication bubbles
Within each bubble there is a replication fork
Bubbles enlarge in both directions & eventually merge/meet, forming duplicate strands
This step uses helicase
Does this by breaking the hydrogen bonds between the nitrogenous bases
Found at the replication fork
Bases are Paired
At the replication fork, enzymes add new complementary nitrogenous bases to parent/original DNA strands
This step uses DNA polymerase
If there is a mismatch the DNA polymerase can backtrack and fix the mistake
There is a challenge with this step DNA polymerase can only add bases (build the complimentary strand) in the 5’ → 3’ direction
No problem for original 3’ strand; the new complementary side starts with 5’ so bases can be added 5’ → 3’
Replication is continuous on this side; known as the leading strand
Bases can be added one after another
The original 5’ strand has the issue… bases cannot be added 3’ → 5"‘ direction
Replication on this side is discontinuous; known has the lagging strand
New complementary bases are added in segments/chunks in the 5’ → 3’ direction
These segments are called Okazaki fragments
Strands Linked Together
This step uses ligase
Each copy has 1 original strand and 1 now strand so it is semi conservative
Replication Fork
Site where the DNA is splitting
Helicase
Enzyme that “unzips”/”unwinds” the DNA
Does this by breaking the hydrogen bonds between the nitrogenous bases
DNA Polymerase
Enzyme that pairs & proofreads the nitrogenous bases
Ligase
Links Okazaki fragment together
Hydrogen bonds reform bonds reform between the new base pairs, leaving 2 identical copies of the original DNA molecule
Each copy has 1 original strand and 1 new strand
Correct DNA Replication Model
Semi - Conservative
What is the instructions to make proteins in DNA based on?
Based on the order of nitrogenous bases
Where are proteins made at
Ribosomes
Ribosomes monomers
Amino Acids
How many types of amino acids
20
The different arrangement of amino acids make…
Different proteins
The order of nitrogenous bases determines…
Which amino acid is added in what order, so ultimately
The order of nucleotides → order of amino acids
The order of amino acids → a specific protein
This process is called Protein Synthesis
RNA
RiboNucleic Acid
One single strand/helix
Found inside and outside the nucleus
RNA made of
Made of nucleotides, but different from DNA
Phosphate
Ribose (Sugar)
Nitrogenous Bases (Adenine, Uracil, Guanine, Cytosine)
Types of RNA
mRNA, tRNA, rRNA
mRNA
Messenger RNA
Copies info from DNA & brings it to ribosome
Located in nucleus, cytoplasm, ribosome
tRNA
Transport RNA
Brings amino acids to ribosome & pairs them with mRNA
Located in cytoplasm and ribosome
rRNA
Ribosome RNA
Directs functions of mRNA & tRNA
Located in ribosome
2 phases of protein synthesis
Transcription
Translation
Transcription
Takes info from DNA and transfers it to a molecule of mRNA
Occurs in the nucleus
Only uses RNA polymerase = enzyme that adds and links complementary RNA nucleotides during transcription
Does the job of helicase, DNA polymerase and ligase all in one
Only uses the leading strand of DNA so new RNA strand can build 5’ → 3’
3 Steps to Transcription
Initiation
RNA polymerase binds to genes promoter region (the start)
Elongation
RNA polymerase unwinds & separates the 2 strands of the DNA helix & adds new RNA bases
This continues until a stop sequence at the end of the gene is reached
Termination
RNA polymerase lands at the stop sequence
mRNA is released and the DNA rezips
In eukaryotic cells, the mRNA has to be modified before it can move to the next phase of protein synthesis
DNA is mixed with 2 types of nucleotide sequence, introns and exons
Process called RNA splicing occurs
Introns are removed/cut out
Exons are stitched together in 1 seamless strand
mRNA is now ready for the next phase of protein synthesis
Base Pairing for RNA
T = A
A = U
C = G & G = C
mRNA bases are added…
in groups of 3
Codons
Sets of 3 mRNA bases that code for an amino acid
Introns
Non-coding segment of nucleotide sequences
Exons
Coding segment of DNA; the parts that will be translated and expressed
Translation
Occurs at the ribosome
During this phase, tRNA carrying amino acids match up with mRNA to interpret the message & build proteins
tRNA is folded into a compact shape
Each tRNA has an anticodon
tRNA anticodons determine the particular amino acid the tRNA carries/transports to ribosome
This specific amino acid will therefore also correspond to the mRNA codon
Ribosomes are made up of what 2 parts
Large subunit and small subunit
mRNA enters and binds on the small subunit
There are 3 slots in the ribosome where the mRNA & tRNA bond and translation occurs
The 3 Slots where mRNA & tRNA Bond
Attachment Site (A Site), Peptide Bond Site (P Site), Exit Site (E Site)
Attachment Site (A Site)
Where tRNA enters ribosome & meets with mRNA
Peptide Bond Site (P Site)
Peptide bonds from between amino acid s of 2 tRNAs
Where the protein chain forms
Exit Site (E Site)
“Naked” tRNA leaves the ribosome & goes off to pick up another corresponding amino acid
Steps of Translation
mRNA leaves the nucleus & enters the cytoplasm
At the same tRNA picks up an amino acid and brings it to the ribosome
The mRNA “start codon”(AUG) enters the A site, then moves into the P Site, signaling the start of the protein
Allows a tRNA carrying amino acid “methionine” to bind to the start codon
Next mRNA codon matches with next tRNA anticodon & amino acid in the A Site
Amino acid from 2nd forms a peptide bond with methionine
New tRNA enters A Site & first tRNA moves into E Site, where it leaves & goes off into cytoplasm to find another corresponding amino acid
Repeat steps 3 - 5 until the end of the mRNA strand is reached
The last code on mRNA is the “stop codon”
Biotechnology
The use of organisms to perform practical tasks for humans
This is accomplished through Genetic Engineering
Through years of experiments, scientists observed bacteria & viruses can swap or absorb new genes from other bacteria & viruses or from their surrounding environment
Recombinant DNA
DNA made from 2 or more different organisms
Plasmids
Many bacteria contain this
Small, circular DNA molecules that are seperate from the main bacterial chromosome
They carry genes & can replicate independently
When a plasmid replicates…
One copy can pass from one generation to another
Results in gene “sharing” among bacteria
This is how antibiotic resistance can form
Can also be used in genetic engineering through gene cloning
Steps to Gene Cloning
Plasmid DNA is removed from a bacteria cell & desired gene is inserted into the plasma
Plasmid is now a combo of original DNA & new DNA - recombinant DNA
Recombinant plasmid is put back in the bacteria cell & replicates, making as many copies as needed
The piece of DNA is cut out of a larger DNA molecule by what enzyme?
Restriction enzymes
Restriction Enzymes
Bacterial enzymes that recognize & bind to short sequences of DNA, then cut the DNA between specific nucleotides within the sequence
Cut the sugar phosphate backbone
Most restriction enzymes make staggered cuts cause…
DNA hanging off the ends of the fragments, known as sticky ends
They are available to bind to any complementary sequence
DNA ligase “pastes” the sticky ends together in the recombinant DNA, repairing the sugar phosphate backbone
The recombinant DNA can now go through gene cloning
One challenge of genetic engineering is…
Finding & isolating the cells that contain the gene of interest
Cells that have taken up the plasmid DNA are identified by growing the bacteria is petri dishes with an an antibiotic
Only the cells that survive the antibiotic exposure have the plasmid DNA
The surviving cells reproduce & form a bacterial colony
Confirmation of the cloned gene can be accomplished by what
Gel electrophoresis
Gel Electrophoresis
A technique used to sort DNA fragments by length
Gel Electrophoresis Steps
DNA from the sample is cut into fragments by
restriction enzymes. The fragments are different sizes
Drops of the sample are placed in wells or slots at one
end of a thin slab of gel (made from agar)
An electric charge / field is applied & the DNA begins to move Because DNA has a negative charge, it migrates toward the positive end of the gel
The DNA separates into fragments according to size, producing a banded pattern. The larger fragments move more slowly & will be closer to the end of the gel with the wells. The smaller fragments are lighter and will travel farther across the gel