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Chapter 7

DNA- major information- carrying molecule, polymer of nucleotides linked by covalent bonds
Transcription- RNA from DNA.
RNA- usually single stranded
mRNA- templates for the production of protein, known as Translation.
Griffith Experiment:
Examined two strains of Streptocccus Pneumoniae. One is S strain and another is R strain. When S strain
was injected to a mice, the mice died. When S is heated, the mice did not die. When R strain in injected,
the mice did not die. However, when R strain and heated S Strain is combined and injected, the mice
died. This is due to the transformation of the R strain from the S Strain.
Avery, Macleod, and McCarty Experiment:
Found out that DNA is the transformation factor.
Hershey- Chase Experiment:
Labelled both proteins and DNA of T2 bacteriophage with sulfur and phosphorous respectively.
Bacteriophage was allowed to inject an E. coli cell and was blended after. It was then brought to a
centrifuge to separate the phage capsids from the infected cells. It was found that the radioactive
phosphorous entered the Cell while the radioactive sulfur did not.
James Watson and Francis Crick- structure of DNA.
DNA- two chains, each chain is a polymer of nucleotides. Each nucleotide is consist of 2-deoxyribose,
phosphate group, and one of four nitrogen containing bases.
Nitrogeneous base are classified as Purine and Pyrimidine.
In DNA, adenine binds with thymine , and guanine binds with cystosine.
The two strands are arranged in an anti parallel fashion and the strands are complementary to each
other.
Bacteria- have single,circular chromosome. It also consists of plasmids, small circular extrachromosomal
pieces of DNA.
Eukarya- have linear DNA chromosomes, in which DNA is complexed with histons, forming nucleosomes.
Archaeons- have single circular chromosome, and has histone like proteins for compaction.

Gene- any segment of DNA that gets transcribed, or copied into a piece of single- stranded RNA, as well
as the associated DNA elements that directs its transcription.
The sugar in RNA is ribose. And Adenine binds with Uracil.
mRNA- coding molecules, translated into proteins.
tRNA- involved in translated, charged with amino acids.
rRNA- structural components of ribosomes.
RNA polymerase- enzyme that transcribes DNA, doesnt need a primer to elongate the RNA
Promoters- where transcription occurs and a site to unwind the DNA.
Template strand of the DNA is use to make an RNA.
In bacteria, sigma factor, transient component of RNA polymerase, binds to the promoter.
Sigma factor- recognition of the promoter region.
In eukarya and archaea, transition factors bind to the promoter and then recruit RNA polymerase.
These binding interactions can be observed with gel shift assay.
Eukaryal mRNAs undergo post transcriptional modifications. A 5 cap and 3 polyAtail are added, introns
are spliced out and exons are join together.
Introns- non protein coding regions.
Translation- information within the mRNA molecules is used to make proteins.Protein Synthesis
Ribosomes- interact with mRNA and read the mRNA sequence, three nucleotides at a time.
Codons- group of three nucleotides
tRNA then comes containing anticodons go complement the codon and bring in the appropriate amino
acids.
In bacteria, translation begins at the first AUG codon after the Shine-Dalgarno sequence.
Shine Dalgarno Sequence- a short sequence in the mRNA recognized by the ribosome.
In eukarya, translation begins at the first AUG after the 5 cap.
As a result bacterial genes may be polycistronic.
While eukaryal genes are monocistronic.
Translation stops when the ribosome reaches a stop codon- UAA,UAG,UGA.
Following translation polypeptides must be correctly folded with the help of Chaperonins.
Post-translational modification such as phosphorylation and glycosyation may occur during protein
processing.
In bacteria, transcription and translation occur almost simultaneously.
In eukarya, two events are separated.
Mutations provide the raw material for evolution.
Base substitution- a mutation in which a single base in the wild type or normal sequence changes.
Missense mutations- mutations that alters a codon.
Silent mutations- mutations does not alter the amino acid sequence of the resulting protein.
Nonsense mutations- converts a normal codon to a stop codon.
Insertion and Deletions or bases also occur. All the codons after an insertion or deletion will be
different. This is called frameshift mutations.
Inversion- sections of DNA are inverted and reinserted into the genome.
Translocation- segment of a chromosome breaks off and reattaches to a different chromosome or a
different location within the same chromosome.
DNA replication
Semiconservative replication- as the double strand DNA molecule unwinds, each strand serves a
template for the formation of a new complementary strand. As a result, two new double stranded
molecules are formed each identical to the original molecule.
oriC- where DNA replications begins in bacteria.
DnaA and Helicase- interact with oriC and initiate replication.
DnaG or Primase- synthesize short segments of RNA needed to prime DNA replication.
Bacteria- DnaA,DnaB,DnaC interact with oriC to help form a replication bubble.
In eukarya, ORC binds to ARS to initiate replication. Recruited Proteins then unwinds the DNA.
In bacteria and eukarya, SSB proteins keeps DNA strands apart.
Elongation
Primase then attaches short segments of RNA to the replication bubble. These are called Primers.
DNA polymerase 3 then extends the primers in a 5 to 3 direction.
Leading strands- elongate continuously.
Lagging strands- elongates discontinuously.
DNA polymerase 1 then removes the primers.
DNA ligase then covalently bonds adjacent nucleotides.
Termination
In circular DNA molecules, replication continues until two replication forks meet.
In linear molecules, telomeres exist on the ends of the molecule, telomerase then completes the
replication process.




Rho independent termination- formation of a hairpin loop followed by a poly U sequence.
Polycistonic- contains the code for multiple peptides.