A plasmid containing the sequence 5??GTCGGATCCTTA?3?3??CAGCCTAGGAAT?5? is combined with BamHI. Write the unpaired bases of the 5??3? sticky end, and then write the unpaired bases of the 3??5? sticky end from left to right as you read them on your screen.
Express your answer as the sequence of the 5??3? sticky end followed by the sequence of the 3??5? sticky end separated by a comma (e.g., AGTGC,TCACG).
A plasmid containing the sequence 5??GTCGGATCCTTA?3?3??CAGCCTAGGAAT?5? is combined with BamHI. Write the unpaired bases of the 5??3? sticky end, and then write the unpaired bases of the 3??5? sticky end from left to right as you read them on your screen.
Express your answer as the sequence of the 5??3? sticky end followed by the sequence of the 3??5? sticky end separated by a comma (e.g., AGTGC,TCACG).
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We have four DNA sequences that we need to digest (cut up), and have 6 restriction enzymes in stock in our lab. We want to find out which restriction enzyme we can use on each sample.
For each sequence, please indicate which restriction enzyme(s) would cut within each sequence. Each sequence may be cut by one, more than one, or no restriction enzymes.
Following the sequences are two questions. Please choose from the same list of enzymes to indicate the correct answer to these questions.
| Restriction enzyme | Digestion pattern |
| EcoRI | 5' G|AATTC 3' 3' CTTAA|G 5' |
| HindIII | 5' A|AGCTT 3' 3' TTCGA|A 5' |
| BamHI | 5' G|GATCC 3' 3' CCTAG|G 5 |
| AluI | 5' AG|CT 3' 3' TC|GA 5' |
| SmaI | 5' CCC|GGG 3' 3' GGG|CCC 5' |
| HbaI | 5' GCG|C 3' 3' C|GCG 5' |
(The | symbol indicates the specific cut sites.)
Each answer may be used once, more than once, or not at all.
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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?
