1. Gene Function: Genes in Action Chapter 11 • Key Knowledge: • molecular genetics • principal events in transcription and translation; cell reproduction: cell cycle, DNA replication, apoptosis; binary fission; gene regulation • prokaryote chromosome and plasmids

2. Gene Action • Gene action involves two processes: • Transcription (t/c) • Translation (t/l) • After the next few slides we will be able to answer the following questions: • How do we get from the bases found in DNA to amino acids? • How do we get from a bunch of amino acids to proteins?

3. DNA Usually double stranded Deoxyribose sugar Bases: C,G A,T RNA Usually single stranded Ribose sugar Bases: C,G,A,U RNA versus DNA Both contain a sugar, phosphate, and base.

4. DNA RNA Protein The Central Dogma:Transcription and Translation Transcription Translation

6. Transcription • Has a number of key processes: • mRNA is made using DNA • mRNA is then modified • During transcription, a DNA sequence is read by RNA polymerase, which produces a complementary, antiparallel mRNA strand.

7. DNA forms base pairs with RNA: DNA RNA C G G C T A A U

8. RNA Polymerase • RNA polymerase is an enzyme complex that catalyses the transcription of DNA to RNA. • It begins transcription by binding to the DNA double helix and gradually unwinds the helix, exposing the DNA template to be transcribed.

9. Transcription Video • Video

10. After Transcription • After transcription occurs you are left with an mRNA strand. • However, this strand needs to be modified further in order to be translated. • This is called post-transcriptional modification. • During this process, the introns in the mRNA strand are removed, and the final mRNA molecule is chemically caped and a poly-A tail is attached.

11. Translation • Involves three types of RNA: mRNA, tRNA and rRNA • Translation involves the use of mRNA (from transcription) held in a ribosome by rRNA (ribosomal RNA) to make a protein. • Amino acids are transferred by the tRNA (transfer RNA) to their matching codons, and hence a protein is assembled.

12. Translation • Translation has three main stages: • Initiation • Elongation • Termination • All three stages occur on the ribosomes • Ribosomes contain 3 binding sites • E - Termination • P - elongation • A - initiation

14. Translation • tRNA brings amino acids to the ribosome

15. tRNA Transfer RNA • amino acid bound to one end • Anticodon on the other end complements mRNA codon

16. tRNA Function • Amino acids must be in the correct order for the protein to function correctly. • tRNA lines up amino acids using mRNA code

17. Reading the DNA code • Every DNA base pairs with a mRNA base • Codon - coding triplet of DNA bases and corresponding mRNA bases • Every group of 3 mRNA bases encodes for a single amino acid

18. Translation Video • Video

19. Reading the mRNA code

20. Reading the mRNA code

21. Synthesising new DNA • DNA needs to be copied exactly so that it can be passed from one cell to another. This process is called DNA replication. • During DNA replication each strand of parental DNA molecule acts as a template strand on which a new strand is synthesised.

22. DNA Replication • DNA replication begins when a region of double-stranded DNA unwinds to form a region of single-stranded DNA. • Single nucleotide are the raw materials for the process of DNA replication. • The nucleotides come into place – where there is a G in the template strand, a C-containing nucleotide is brought into place. • In this way, the base sequence of the double-strand of DNA controls the order of the nucleotides in two new single strands of DNA. • This process is catalysed by the enzyme DNA polymerase. • Each nucleotide joins to its neighbour in the chain by a strong sugar-phosphate bond.

23. The strand is always copied from 5’ to 3’.

24. Which model of DNA replication? How do we know which model is correct?

25. The mystery of the model of DNA replication was solved in 1958 by Meselsohn and Stahl. • They took bacterial cells and allowed them to multiply for several generations in a growth medium containing ‘heavy’ nitrogen (15N) only, so that all nitrogen atoms in these bacterial DNA were heavy nitrogen. • These bacterial cells were then transferred to a new growth medium containing only normal nitrogen (14N). • After one generation, a sample of the new bacterial cells was collected. Any new DNA made by this new generation could only include 14N atoms. • The DNA was extracted and the relative weight of the DNA was determined using a centrifuge. • The lower down the tube, the heavier the DNA.

26. They obtained a single band with their experiment. Showing that replicated DNA contained one original chain and one new chain. This means that DNA replication is semi-conservative

27. apoptosis; binary fission; gene regulation, prokaryote chromosome.