Problem Set 2 ?You used the sequence from #4 as a probe to clone the gene respon

Question

Problem Set 2 ?You used the sequence from #4 as a probe to clone the gene responsible for liver cel degradation from a genomic library, you sequenced the clone and obtained the following DNA sequence. Answer the questions regarding the following DNA sequence: 5' 110 120 130 140 G-A-A-T-T-c-T-C-T-T-C-C-G-G-A-A-G-A-T-C-G-A-G-T-G-A-T-A-T-A-A-G-C-T-T-GC-A-G-G-A-G-T-G-A-T-G-C 150 A-T-G-G-C-c-T-A-T-C-C-G-T-T-G-C-T-T-C-C-G-G-A-T-C-G-A-G-T-A-A-T-T-T-T-A-A-G-C-T-T-T-G-G-G-A-A-A 160 170 188 198 1100 T-C-GA-G-G-A-A-T-T-C GIVEN: () RNA polymerase binds at a T-A-T-A sequence and STARTS transcription at the 7th base from the binding site. (i) RNA polymerase terminates immediately before a T-TTGGG-A-A-A sequence. (iii) Intron ends have the sequences cocc and CCGG regions (they come together to form a loop). (iv) The RNA is cut at the 3' end of a A-UU-u-u sequence by the POLY-A enzyme complex and a poly A tail is added to the cut end. (o) The RIBOSOME binding site is CA-GG-A-G (vi) All eukaryotic proteins START with the amino acid, methionine (met) a. b. c. What is the sequence of the primary RNA? What is the exact sequence of the mature mRNA? What is the sequence of the liver cell degradation protein? (use the genetic code in your book)

Explanation / Answer

RNA polymerase recognizes consensus TATA sequence (TATAAT). However, there is no such sequence in the given sequence. So, I assume TATAAG that starts from 27th position as TATA sequence. This TATA sequence ends at 32nd position. Given that transcription starts at 7th postion from TATA sequence, i.e, 39th position (G).

Also given that polymerase terminates before TTTGGGAAA sequence, which is at 88th position. Therefore, transcription occurs from 39th nucleotide to 87th nucleotide.

a.Sequence of primary RNA:

GGA GUG AUG CAU GGC CUA UCC GUU GCU UCC GGA UCG AGU AAU UUU AAG C

Given that introns ends have GGCC and CCGG

b. sequence of mature mRNA:

GGA GUG AUG CAU AUC GAG UAA UUU UAA GC

c. sequence of protein starting from AUG:

Met His Ile Glu stop

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