Dr Michelle Smith BIO350 Spring 201 7 Open Reading Frame Act

Dr. Michelle Smith

BIO350

Spring 201

7

Open Reading Frame

Activity

Craig Venterand his colleagues wanted to know if a complete genetic system could be producedusing chemicals and a computer generated DNA sequence. In 2010, an article was published describing the first self-replicatingsynthetic bacteria (M. mycoides JCVI-syn1.0)that has acomputer sequence as its parent. To verify that they had synthesized a new organism, Venter’steam put DNA sequences inM. mycoides JCVI-syn1.0 that spell out secret messages.

The same DNA sequence is printed out 3 times (just broken into different triplet reading frames). Use theSINGLE LETTER AMINO ACID sequence abbreviations (hint, hint) in the codon table to uncover the secret message. The Open Reading Frame (ORF) starts with the codon 5’ ATG 3’ (which is written as5’ AUG 3’ in the codon table because mRNA is translated into amino acids). The DNA sequencebelow does not encode any introns.

Watson

#1 5’- AAA TTT ATC GTT CTG TGT TCT CTA CTC CAA TTG CAC GAC ACA TTG AA -3’

3’- TTT AAA TAG CAA GAC ACA AGA GAT GAG GTT AAC GTG CTG TGT AAC TT -5’ #4

Crick

Watson

#2 5’- A AAT TTA TCG TTC TGT GTT CTC TAC TCC AAT TGC ACG ACA CAT TGA A -3’

3’- T TTA AAT AGC AAG ACA CAA GAG ATG AGG TTA ACG TGC TGT GTA ACT T -5’#5

Crick

Watson

#3 5’- AA ATT TAT CGT TCT GTG TTC TCT ACT CCA ATT GCA CGA CAC ATT GAA -3’

3’- TT TAA ATA GCA AGA CAC AAG AGA TGA GGT TAA CGT GCT GTG TAA CTT -5’ #6

Crick

1. What is the secret

message and where?

UUU, UUC- F

UUA, UUG- L

UCU, UCC , UCA, UCG- S

UAU, UAC- Y

UAA, UAG-Stop codon

UGU, UGC- C

UGA- Stop codon

UGG- W

CUU, CUC CUA CUG- L

CCU CCC CCA CCG- P

CAU CAC- H

CAA CAG-Q

CGU CGC CGA CGG- R

AUU AUC AUA - I

AUG- M

ACU ACC ACA ACG- T

AAU AAC- N

AAA AAG- K

AGU AGC - S

AGA AGG- R

GUU GUC GUA GUG- V

GCU GCC GCA GCG - A

GAU GAC - D

GAA GAG- E

GGU GGC GGA GGG- G

Solution

sequence 1 template: 5’- AAA TTT ATC GTT CTG TGT TCT CTA CTC CAA TTG CAC GAC ACA TTG AA -3’

mRNA:                   3\' - UUU AAA UAG CAA GAC ACA AGA GAU GAG GUU AAC GUG CUG UGU AAG UU - 5\'

Nontemplate strand: 3’- TTT AAA TAG CAA GAC ACA AGA GAT GAG GTT AAC GTG CTG TGT AAC TT -5\'

mRNA :                5\' - AAA UUU AUC GUU CUG UGU UCU CUA CUC CAA UUG CAC GAC ACA UUG AA -3\'

sequence 2 template: 5’- A AAT TTA TCG TTC TGT GTT CTC TAC TCC AAT TGC ACG ACA CAT TGA A -3’

mRNA:                     3\'- U UUA AAU AGC AAG ACA CAA GAG AUG AGG UUA ACG UGC UGU GUA ACU U -5\'

Amino acid sequence:                                                             M      R     L         T     C      C    V     T

Nontemplate strand:   3’- T TTA AAT AGC AAG ACA CAA GAG ATG AGG TTA ACG TGC TGT GTA ACT T -5’

mRNA:                  5\' - A AAU UUA UCG UUC UGU GUU CUC UAC UCC AAU UGC ACG ACA CAU UGA A - 3\'

sequence 3 template : 5’- AA ATT TAT CGT TCT GTG TTC TCT ACT CCA ATT GCA CGA CAC ATT GAA -3’

mRNA:                   3\' - UU UAA AUA GCA AGA CAC AAG AGA UGA GGU UAA CGU GCU GUG UAA CUU - 5\'

Nontemplate strand: 3’- TT TAA ATA GCA AGA CAC AAG AGA TGA GGT TAA CGT GCT GTG TAA CTT -5’

mRNA:                  5\' - AA AUU UAU CGU UCU GUG UUC UCU ACU CCA AUU GCA CGA CAC AUU GAA - 3\'

Secret message in DNA template strand 2: 5\' - TAC TCC AAT TGC ACG ACA CAT TGA - 3\'

                                              mRNA:   3\' - AUG AGG UUA ACG UGC UGU GUA ACU - 5\'

Amino acid sequence:                                M      R     L         T     C      C    V     T

The secret message is hidden in the template starnd of sequence 2 or ORF 2.