You are studying an integral membrane protein of the plasma membrane. It has a single transmembrane domain and the N-terminal portion of this protein is extracellular. You decide to perform mutagenesis studies of the stop-transfer sequence, and you mutate all of the valine and isoleucine residues within this sequence to lysine and arginine residues. What do you predict will happen to the transport of this protein and why (be detailed in your answer)?
You are studying an integral membrane protein of the plasma membrane. It has a single transmembrane domain and the N-terminal portion of this protein is extracellular. You decide to perform mutagenesis studies of the stop-transfer sequence, and you mutate all of the valine and isoleucine residues within this sequence to lysine and arginine residues. What do you predict will happen to the transport of this protein and why (be detailed in your answer)?
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SEQUANCE:
5'- GCU UGG GCU GCU GAU GCU CUU UAA CAU AGA GCU AAU AUU -3'
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3- Convert your mRNA sequence into a DNA sequence, then make itdouble-stranded by writing the complementary sequence below.
4- Transcribe (write an RNA sequence) and translate (write anamino acid sequence) both the "top" and "bottom" strands startingwith the first nucleotide. You must use the genetic code in yourtextbook or in your lecture notes for this step. DO NOT use thetable above. If you encounter a stop codon, your protein sequenceis done at that point.
Use the single letter designation for amino acids as givenbelow.
Alanine (A) | Aspartic acid (D) | Asparagine (N) | Arginine (R) |
Cysteine (C) | Glycine (G) | Glutamic acid (E) | Glutamine (Q) |
Histidine (H) | Isoleucine (I) | Leucine (L) | Lysine (K) |
Methionine (M) | Proline (P) | Phenylalanine (F) | Serine (S) |
Threonine (T) | Tryptophan (W) | Tyrosine (Y) | Valine (V) |
5- Use your original DNA sequence from #3. Add an A to the 5'end of the "top" strand, then write the complementary sequence onthe bottom. Add a GC to the 5' end of the
"bottom" strand, then write the complementary sequence on the top.Transcribe and translate both the "top" and "bottom" with this newsequence.
What kind of mutation(s) was introduced in the "top" strand?
What kind of mutation(s) was introduced in the "bottom"strand?
6- Use your original DNA sequence from #3. Starting at the 5'end on the "top" strand of your original sequence from part 3,delete the second A, the third G, and the fourth C along with thecomplementary nucleotides on the bottom strand. Transcribe andtranslate both the "top" and "bottom" with this new sequence.
What kind of mutation(s) was introduced in the "top" strand?
What kind of mutation(s) was introduced in the "bottom"strand?
7. Use your original DNA sequence from #3. Starting at the 5'end of the "top" strand, change the third G to an A, then changethe complementary nucleotide in the bottom. Starting at the 5' endof the "bottom" strand, change the third C to a T, then change thecomplementary nucleotide on the top. Transcribe and translate boththe "top" and "bottom" with this new sequence.
a. What kind of mutation(s) was introduced in the "top"strand?
b. Whatkind of mutation(s) was introduced in the "bottom" strand?
