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Cystic Fibrosis From Gene to Protein Class instructions In this lesson • Look at the normal and mutated CFTR gene • Make messenger RNA (mRNA) from DNAtranscription • Make protein from mRNA- translation The devil is in the detail! • The 5’ prime and 3’ prime ends of the bases must be round the right way! IMPORTANT: Do not take bases apart!!! Correct base pairing is critical! • Green (Guanine) pairs with yellow (Cytosine) • Blue (Adenine) pairs with orange (Thymine) Lesson – part 1 Normal CFTR Gene ΔF508 CF Mutation Sense Sense Antisense Antisense Transcription – Step 1 Base pairing in mRNA- use brown sugars • Green (Guanine) pairs with yellow (Cytosine) • Blue (Adenine) pairs with light blue (Uracil) Transcription – Step 2 • To make an mRNA, you must use the antisense (red sugars) strand as as the template as base pairing is complimentary. Resulting mRNA Normal CFTR mRNA ΔF508 CFTR mRNA Translation • Turns mRNA into protein • Use transfer RNA (tRNA) which are attached to amino acids as intermediate • Amino acids have a direction too! N-terminal C-terminal Transfer RNA UAA Isoleucine CAA Valine CCA Glycine AAA Phenylalanin e UAA Isoleucine tRNA – holds an anticodon at one end, and an amino acid at the other Using mRNA in 5’-3’ orientation, translate into protein. Separate tRNA & protein from mRNA Resulting protein Normal CFTR protein Isoleucine – Phenylalanine Glycine ΔF508 CFTR protein Isoleucine – Glycine Valine Polymerase Chain Reaction • Using the original chromosomes, separate strands • Find where primers stick to each strand Genetic Test • Using the original chromosomes, see what a PCR product will look like. • Remember: Each individual will have 2 chromosomes - one inherited from mother and one from father. DNA gel genetic test result