Research Presentation C. McLemore

Identification and Isolation
of Stress Related Gene in
Gray (Eschrichtius
robustus) and Fin Whales
(Balaenoptera physalus)

Christin McLemore
June 04, 2009
School of Aquatic and Fishery Science
University of Washington
Box 355020
Seattle, WA 98195

Background
• North Pacific is home to over 80 species of cetaceans
• Whales
• Dolphins
• Porpoises

• OrderCetacea
• Sub-Order Odontoceti
• Sub-Order Mysticeti

Gray Whale, Eschrichtius robustus

• Females are typically 11.7 and 15.2 m in length and males are 11.1 to 14.3
m (Evans 1987).

• Grey colored skin with barnacles (Cryptolepas rachianecti) and barnacle
scars

• Only large whale where the upper jaw extends over the lower jaw

(Reeves, 1987)

Fin Whale, Balaenoptera physalus

• Males and Females are typically 18 to 21 m, up to 27 m

• Clearly defined grey, white, brown and black, chevron-shaped coloration pattern on
the right side, behind the head and an asymmetrical coloration of the lower jaw:
white on right, dark on left

• The status of Atlantic and Pacific Fin Whale assigned by the Committee on the
Status of Endangered Wildlife in Canada (COSEWIC)
• April 1987 – “of special concern”
• May 2002 – Pacific Fin whale is classified as THREATENED

Environmental Factors
• Pollutant
• organochlorine compounds like
DDTs, hexachlorocyclohexanes
(HCHs), chlordane compounds (CHLs) and
polychlorinated biphenyls (PCBs) can disrupt
normal endocrine physiology in marine mammals
(Tanabe, 2002).
• Climate Change
• reduction of available habitat
•the acidification of the oceans as they absorb
growing quantities of CO2

• Disease
• Increased susceptibility of cetaceans to diseases
• reduced reproductive success
•body condition
•survival rates

(World Wide Fund, 2007)

Objectives
• Identification of genes of interest
•Glutathione S-transferase A4 (GStra)
• Superoxide dismutase [Cu-Zn] (SD)
• Heat shock 70kDa protein 6 (HSP6)
• Interleukin-2 precursor (IL-2)
• Interleukin 17 (IL17)
• Interleukin 13 precursor (IL13)
• Toll-like receptor 5 (TLR5)
• Cyclooxygenase (COX-2)

• Isolation of the expressed gene of interest
• Interleukin 17
• Prostaglandin-endoperoxide synthetase-2
• tRNA synthetase
• Glutathione S-transferase A4

• Homology of genes from other organisms with Grey and Fin Whale

Results

Figure 1: Row one: Lane (1)100 bp ladder, (2) Figure 2: Lane (1) 100 bp ladder, (2) Negative
Negative control Gstra, (3) Grey whale Gstra, (4) control IL17, (3) Grey whale IL17, (4) Fin whale
Fin whale Gstra, (5) Negative control HSP6, (6) IL17, (5) Negative control IL13, (6) Grey whale
Grey whale HSP6, (7) Fin whale HSP6, (8) Negative IL13, (7) Fin whale IL13, (8) Negative control
control SD, (9) Grey whale SD, (10) Fin whale SD. IL2, (9) Grey whale IL2, (10) Fin whale IL2, (11)
Row 2: Lane (1) 100 bp ladder, (2) Negative control Negative control TLR5, (12) Grey whale
COX2, (3) Grey whale COX2, (4) Fin whale COX2. TLR5, (13) Fin whale TLR5, (14) 100 bp ladder.

Reference Grey Whale
Reference Taxa Fin Whale Coverage
Size Coverage

Homo sapien
Interleukin 17 (AY460616)
7,422 1,729 - 3,093 2,385 – 2,756

Prostaglandin-
Homo sapien
endoperoxide 11,449 4,041 – 5,563
(AY229989)
synthetase-2

Bos taurus
tRNA synthetase (BC122671)
1,538 1 - 360

Glutathione S- Homo sapien
21,305 12,394 – 12,942 12,790 - 13,423
transferase A4 (AY878121)

Interleukin 17
Eschrichtius robustus Balaenoptera physalus

CTCCAGATCACAGAGGGGTACCTCTCAGGGTCCTCATTGCGGCTATAAGG AGGATAAATCTCTGTATTCTTCTACCCTGTATGTATCAGACTGCCAATTA
GAGACAAAAGCCAGGAAGAGAGTGAAATCCTCGTAGAGCAGGAAGGAAAT AATCACCTTGAGAATGGTATTACTCACTCAGAATCATCCGTCAAAACTGG
CAAACAGGAATAAACAAAGGGAGACAAATTCACAAGGGTGCAAATGATAG AAGAGCCATTGTGTAAACCAATGATTCTCAAACTTTCATTCTACAGATTT
GCATTGTGAATTCTCTGAGTGCTGTTAGTCTGAAAAAGAAGTAGTTTGGG CTTTGATAGAGCACAGCTCACCCCACCCCCCATAACACCTATTCTACTAG
GAAAACTTGTGTTTGCAAGCCCATGAAGTGTTATAGAAAAAAAGACAATG TTCTGATTGTCCCAGAAATCCATCCAAACGAACGGTGGTCATCATTCTCA
ATTAGTTTTTCCCAATTTCTTTCCAACATACACAAAAGAAGATAGACTGA TATTTATGACACCCCAAGTCTGTTGTTTATTTATTTTAAAAAGAAATAGA
AACTACAGAATGGATTTAAGTTTGACACAAAGGTTAATTTGTTTAGGGTA CATTTTTCATAGGAGATTAGTGCTTGAATTCTCTTCACCAGAGCTCCCAA
CATTTGATGGATGAAAAAAGACTAAAGGAATAGTATAGAAACAAGATAAC AGAAGATTTGTTGAAAATAACTATTAATTAACTGGGATTTTTGTGCTTTT
TGTTAATCTGACTTAATGAAGTCTGTCTTTGAAACAGAGAATGCACCCAA TTGGTTGTATTAACACTGGGAAAATATAGCTTGAATACCAGTGGAAGAAG
ATTTTATCTGACAGCACGCTGATGGAACAGAGCCCACGGCTGATATGAAG TATCATGCTGGGTATTTCCACCCTTTTTTGACTTAGACAACATCCCCAAC
GGCTGCAAACAAAATCTTAAGCCCCTTTATAACCTTCTCTTCTCACTCCC AGACCAATATTTGCTGTGTCCTTAGTACATCTAGTATTTAGABNNATNAA
CTCCAGCTGAGTGAATAATTTGCTTCGACTTCTGAATTAATTTATTTGAA GGGTTAAATACNAAATGTATTTTGATAATTTGATTTTAGGCAAATAANNN
CTATTCTTAGTAAGTAGAAGTAAGAAGACAAGAAACCCTACAGTCAGGTG NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN
AATCTGTAGGCTCCATATAACTGAGGTTTATTTGCCTAAAATCAAATTAT NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN
CAAAATACATTTTGTACTTAATCCATCTAAATACTAGATGTACTAAGGAC NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN
ACAGCAAANTATTGKTCTGTTGGGGATGTTGTCTAAGTCAAAAAAGGGTG NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN
GAAATACCCAGCATGATACTTCTTCCACTGGTATTCAAGCTATATTTTCC NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN
CAGTGTTAATACAATCAAAAAAGCACAAAAATCCCAGTTAATTAATAGTT NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN
ATTTTCAACAAATCTTCTCTGGGAGCTCTGGTAAAGAGAATTCAAGCACT NNNNNNNNNNNNNNNNNNNNNNNNNNNTCCATCAAATGTACCCTAAACAA
AATCTCCTACGATAAATGTCTATTTCTTTTAAAAATAAATAAACAACAGA ATTAACCTTTGTGTCAAATTTAAATCCATTCTGTAGTTTCAGTCTATCTT
CTTGGGGTGTCATAAATATGAGAATGATGACCACCGTTCATTTGGATGGA CTTTTGTGTATGTTGGAAAGAAATCGGGAAAAACTAATCATTGTCTTTTT
TTTCTGGGACAATGAGAACTAGTAGAATAGGTGTTATGGGGGGTGGGGTG TTCTATAACACTTCATGGGCTTTGAAACACAAGTTTTCCCCAAACTACTT
AGCTGTGCTCTATCAAAGAAATCTGTAGAATGAAAGTTTGAGAATCATTG CTTTTTCAGACTAACAGCACTTAGAGAATTCACAATGCCTATCATTTGCG
GTTTACACAATGGCCCTTCCAGTTTTGACGGATGATTCTGAGTGAGTAAC CCCCTGTGAATTTGTCTCCCTTTGTTTATTCCTGTTTGATTTCCTTCCTG
ACCATTCTCAAGGTGATTTAACTGGCAGTCTGATACATACAGGGTAGAAG CTCTACGAGGATTTCACTCTCTTCCTGGCTTTTGTCTCCCTTGTAGACGC
AATACTGAGATTATCCT TAATGAGGACCCTGAGAGGTACCCCTCTGTGATCTGG

Reference Grey Whale
Reference Taxa Fin Whale Coverage
Size Coverage

Homo sapien
Interleukin 17 (AY460616)
7,422 1,729 - 3,093 2,385 – 2,756

Prostaglandin-
Homo sapien
endoperoxide 11,449 4,041 – 5,563
(AY229989)
synthetase-2

Bos taurus
tRNA syntetase (BC122671)
1,538 1 - 360

Glutathione S- Homo sapien
21,305 12,394 – 12,942 12,790 - 13,423
transferase A4 (AY878121)

Prostaglandin-endoperoxide synthetase-2
Eschrichtius robustus
GTGTGAAGGTGAGTGAGAGGAAGCAATTAGACATGTATTAACTGTAACTG
AGATTGGATTGCTACCTAGAAAATTTGACCATAAACCATGATTTATTTGT
TCATAAAGCCATTTTTTCCTTATGGATATCTCTGGAGATTTTTTGAAAGT
GAGTCCATGTAAACTACTCCATGAAAGTGAATGGGCATTTTGGTATAATG
GCATGATGTTTTCAAATTCTATCATTAAAACTGAATTAATGCTGTAAACA
AAACANNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN
NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN
NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNTCTAGGT
GGACTTAAATCATGTTTATGGTGAATCTTTGGAGAGACAGCATAAACTGC
GCCTTTTCAAGGATGGAAAAATGAAATATCAGGTTTGTCCCATTTAAATA
TTAAGAATTAGTTATGACTCACCACCCACCTATGTTCAAAATCTCTCATG
ATTAAAATTTGTTCCTTACCTTTTT

tRNA syntetase


CTGTGGGAATTAATTGGTGCTAAGGTCTAACATTGTTCTAATCTGAGTGC
CAGGGAATGTTTTCTGTCCCTTCTCACCCCTACATGCTTGTGGTATTTTT
TTTTTATCTTAGGGAATTCCCTGGCASGACCGTGGCTAAGGCTCTGCGCT
TTCACTGCCAAGGGTGCAAGTTAAATCCCTGGTCAGGGAACTAAAATCCC
ACAAGCTGTGCTGTGTGTCCCCCGCCCCCCCCCCCCAAAAAAAAAAAAAA
AACCYCAGGCATTTTTAATTTTTTGGGGCCCAACCCCTTAGGGGACCCCG
AATCCATCTCAGGACCTTTGCACTTACTAGTTCTGCTTGAAACCCTCTTT
CCTCAATCCTGTAATGGATCCCGATTT

Glutatione S-transferase A4
TGACCCCGTAANACCTGCAGGGTATAAACATTTTACATTGGTTATAATAC
TTGAGATGGAGAAAATGATGGAGCTGTGGAATCAGTGCAGTATCTGAGAA
GGGCTGAAACGACGACCTTGTCCTTTTATCCTTTTACGAATAACATCAAC
TAAGAAGAAGTTCCCTATATGAGAGGGAGAGAGCGAAAAAGGTGTAGAAA
TGAAAGCATCCTTAAGTATCTGCTTTACATCAGAATGAGACCCTAAAAAG
GGGAAGGAAATATTAATTCTAGATTTAGCATCCAGGGCTTTAAAAGGATA
CCTCTATTCCATAAATGTATCTACAGAAAAGAATTAGCCCTGATGCATTC
AGGGGAAGTGATGTAAGACGTAAAGCCACTGGTCCTGTCCAAGGAGTCTA
ATCTCATTTGCCAGTTCTGTGTTTCTGTGTGTCAGGACTCACAGTCATAA
ACTAGACATGACAGTAAAAAAGCAGGCTTCCTTTCACGTCTTCTAAGACA
GGGAAAATTAAA

Balaenoptera physalus
AATWAGATACTTTCCTGTGTTCGANAAAGGTAGGTGGCAGTGAAGCCTGT
GAACGCGGCATCACCAGACCACAAACGCCCCTTTCTGCTCAGGCAGAGTA
AAGCTGAGCCCTGCTAGACCAACCTCCTGAGGGCTCACAGGTGCCATCAG
CACAAAGCCAGGATGTCTTGGAGCCGCCCAGACTCTTCCAGAGTCTTCCA
TGTTTTCTGGGAAAAATATGGGCTCTCCCACGTGTGCATTGTCATTGCTT
ACATTCGCCAGGTGCCACCTTGTGCAGGGTACAGAAGAGAAGAAGCCACC
TACTTCATTCATTCGTTCATCATCCTACCTGATAGTTTGGTATAAAGAGA
AGAACCATTGCTGTTTTATTTCTCTTCCCTGTGCTCTTTTTGTTTTCTCT
TTTGTTACTTCTTGTCTCATTGCCGTATCTGTTTTTCTGAGCTTCAAACT
CCTCCTCTCTTTTCCTCAACTGAGAGGGACTATTTTAAATGCTTCAATTA
AAAAACAAGAGAACGTGAAAATCTGGAATTGGCAGAC

Prostaglandin-endoperoxide synthetase-2

Acknowledgement
Dr. Steven Roberts

Research Scientist
Sam White

National Oceanic and Atmospheric Association
Seattle, Washington
Jina Ylitalo

Capstone Coordinator
Greg Jensen

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Research Presentation C. McLemore

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