Molecular Evidence for Evolution Name:

Pre Lab Activity:Genes code for amino acids, amino acids code for proteins and proteins build body structures. Therefore, one way to observe the relatedness of species is to examine their genetic sequences. Generally, the more similarity that exists in these species, the more likely species are to be related to each other.The amino acids below are found in the protein beta hemoglobin. This protein is generally 146 amino acids in length. To conserve space, only the amino acids where differences exist have been provided to you.

Amino Acid Number

Species 1 Species 2 Species 3 Species 4 Species 5

1 Valine Valine Threonine Valine Valine2 Histidine Histidine Leucine Histidine Histidine4 Threonine Threonine Serine Threonine Threonine5 Proline Proline Alanine Proline Glycine8 Lysine Lysine Aspartic Acid Lysine Lysine9 Serine Asparagine Alanine Serine Serine

10 Alanine Alanine Histidine Alanine Alanine12 Threonine Threonine Threonine Threonine Alanine13 Alanine Threonine Serine Alanine Alanine21 Aspartic Acid Aspartic Acid Glutamic Acid Aspartic Acid Aspartic Acid22 Glutamic Acid Glutamic Acid Lysine Glutamic Acid Glutamic Acid33 Valine Leucine Valine Valine Valine50 Threonine Serine Serine Threonine Threonine52 Aspartic Acid Aspartic Acid Serine Aspartic Acid Aspartic Acid56 Glycine Glycine Serine Glycine Serine69 Glycine Glycine Serine Glycine Glycine73 Aspartic Acid Aspartic Acid Glutamic Acid Aspartic Acid Aspartic Acid76 Alanine Asparagine Histidine Alanine Alanine87 Threonine Glutamine Glutamine Threonine Glutamine104 Argenine Lysine Lysine Lysine Argenine111 Valine Valine Serine Valine Valine112 Cysteine Cysteine Alanine Cysteine Cysteine113 Valine Valine Glutamic Acid Valine Valine114 Leucine Leucine Serine Leucine Leucine115 Alanine Alanine Glutamic Acid Alanine Alanine116 Histidine Histidine Leucine Histidine Histidine120 Lysine Lysine Histidine Lysine Lysine121 Glycine Glycine Aspartic Acid Glycine Glycine122 Phenylalanine Phenylalanine Lysine Phenylalanine Phenylalanine123 Threonine Threonine Serine Threonine Threonine125 Proline Glutamine Alanine Proline Glutamine126 Valine Valine Valine Valine Leucine130 Tyrosine Tyrosine Phenylalanine Tyrosine Tyrosine

The old way of doing things was to count the differences between amino acids by hand and develop a phylogenic tree. For fun, compare the differences between species 1 and the other species and record it on the chart! Use the provided highlighter to count the number of differences that exist between species 1 and the other species. Record this information in the chart below.

Number of Differences from Species 1

Species 2Species 3Species 4Species 5

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This method, while accurate and effective, is rather time consuming. It also only allows us to compare organisms to each other one at a time….

Lab Activity:

The new method of doing things is to utilize the vast amount of information available on the Internet.Both of the sites you will use today access information available on the National Center for Biotechnology Information website.

National Center for Biotechnology Information (http://www.ncbi.nlm.nih.gov)

The National Center for Biotechnology Information is a website sponsored by the federal government. The goal of the site is to advance science and health by providing access to biomedical and genomic information.

Pre-Lab Questions:

1. In today’s activity you will compare the gene sequences between whales and different mammals. Two of the mammal groups you will compare whales to are the orders known as artiodactyls and perissodactyls.

In our analysis today, we will look at the protein known as cytochrome b. What is the function of cytochrome b?

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When comparing species such as whales and other mammals, why do you think we use a protein like cytochrome b?

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Go to Google or some other Internet search engine. Next to each term, identify the key characteristic of each group-artiodactyls and perissodactyls. Then, in the spaces below each group, identify two examples of artiodactyls and two examples of perissodactyls.

Artiodactyls (define): Perissodactyls(define):

Examples: Examples:

1. 1.

2. 2.

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Part I – Finding Sequences

1. Open the Internet and type the NCBI homepage into your browser.

(http://www.ncbi.nlm.nih.gov)

2. The aim of our investigation today is to compare the cytochrome b sequences among different mammal groups. In the box next to “Search,” click on the dropdown menu and select the “Protein” Database. In the box next to “for” type the phrase “cytb Mammalia” and click on the “Search” button (cytb is one of the server’s abbreviations for cytochrome b).

3. Clicking on this page takes you to the results page.

Each result lists a hyperlink to the protein as well as the scientific, Latin name of the organism from which the protein was obtained. As you scroll through the results, you will notice that there are upwards of 2,000 pages of entries for this particular protein.

Approximately how many amino acids make up a complete, mammalian cytochrome b protein?

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RIGHT HERE!

4. As you click through the result pages, you may notice there are proteins that are fragments and are much smaller in size than the complete proteins. You want to be careful NOT to select these organisms in your investigation. All protein samples you investigate should be the full size and listed as CYTB proteins.

5. To access each individual organism, you must click on the hyperlink for the organism.

The first organism to come up in this example is Herpestes javanicus. Clicking on the blue hyperlink tells us that this organism is commonly known as:

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6. This organism page has a great deal of information. In addition to the organism’s taxonomy, it provides links to research conducted on the protein, and, important for our investigation, a copy of the protein sequence for the organism is found at the bottom of the page.

Top of page Bottom of page

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7. The mongoose, while an interesting subject for investigation, is not a species that we want to investigate in full detail today. The topic of our investigation, you may recall is to focus on the evolution of whales. Click “Back” in your browser to return to the mammal sequence page.

Once on the page, you can narrow the search even further by searching the protein database for “Cetacea,” (the mammalian order to which whales belong).

8. The first hyperlink is for the whale, Balaenoptera musculus, commonly known as the blue whale.

The information provided is organized in the exact same way as the page you viewed earlier. Our interest is in the amino acid sequence information. Although, as you saw earlier, the amino acid sequence is listed at the bottom of the page, this format will be difficult for us to deal with when we conduct our analysis. The easier format can be found by clicking on the “FASTA” link found at the top of the page.

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Taxonomic Information

Protein Sequence

9. On the FASTA sequence page, use your mouse to select and copy the sequence. This is shown for you below.

10. After you select and copy the information, open a Microsoft Word document and paste this information into your Word document.

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Click on this “FASTA” link

11.Save your Word Document and modify your sequences so that they can be interpreted. To do so, you must remove any blank spaces from the headings so that the computer does not mistake letters for amino acids. In other words…..

The sequence given for the blue whale in FASTA format looks like this:

>gi|5835008|ref|NP_007068.1|CYTB_10457 cytochrome b [Balaenoptera musculus]MTNIRKTHPLMKIINDAFIDLPTPSNISSWWNFGSLLGLCLIVQILTGLFLAMHYTPDTMTAFSSVTHICRDVNYGWVIRYLHANGASMFFICLYAHMGRGLYYGSHAFRETWNIGVILLFTVMATAFVGYVLPWGQMSFWGATVITNLLSAIPYIGTTLVEWIWGGFSVDKATLTRFFAFHFILPFIIMALAIVHLIFLHETGSNNPTGIPSDMDKIPFHPYYTIKDILGALLLILTLLMLTLFAPDLLGDPDNYTPANPLSTPAHIKPEWYFLFAYAILRSIPNKLGGVLALLLSILVLALIPMLHTSKQRSMMFRPFSQFLFWVLVADLLTLTWIGGQPVEHPYVIVGQLASILYFLLILVLMPVTSLIENKLMKW

If you tried to enter this into a sequencing program, all the letters would be interpreted as amino acids, and you would not be able to accurately compare species. To avoid this problem, you must modify the information you obtained. Thus, the blue whale should be entered as:

>Blue.whaleMTNIRKTHPLMKIINDAFIDLPTPSNISSWWNFGSLLGLCLIVQILTGLFLAMHYTPDTMTAFSSVTHICRDVNYGWVIRYLHANGASMFFICLYAHMGRGLYYGSHAFRETWNIGVILLFTVMATAFVGYVLPWGQMSFWGATVITNLLSAIPYIGTTLVEWIWGGFSVDKATLTRFFAFHFILPFIIMALAIVHLIFLHETGSNNPTGIPSDMDKIPFHPYYTIKDILGALLLILTLLMLTLFAPDLLGDPDNYTPANPLSTPAHIKPEWYFLFAYAILRSIPNKLGGVLALLLSILVLALIPMLHTSKQRSMMFRPFSQFLFWVLVADLLTLTWIGGQPVEHPYVIVGQLASILYFLLILVLMPVTSLIENKLMKW

The greater than (>) symbol indicates that what comes next is a name. This means for a two or more word named organism, you must put a period in between words in the species name. For example, if you sequence a Blue Whale, you must modify its name to read as:

>Blue.Whale

If you fail to do so, the program will begin to read the w-h-a-l-e (in whale)… as amino acids!!!

12.Try out the sequencing for yourself. Use the program to find cytochrome b sequences for the following whales, marine mammals and possible land relations of whales. Find each organism’s sequence by searching the protein database using the abbreviation of the protein (cytb) and the scientific name of the organism in quotation marks.

Be sure to double check your spelling for each scientific name, and make sure that you select a full copy of the cytochrome b protein (it should be approximately 378 amino acids in length).

Organism Common Name Scientific Name Order Relationship to

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Enter: cytb and the “Scientific Name” of the organism in this box.

WhalesWhale 1 Blue Whale Balaenoptera musculus Cetacea -Whale 2 Humpback Whale Megaptera novaeangliae Cetacea -

Whale 3 Sperm Whale Physeter catodon Cetacea -Whale 4 Killer Whale Orcinus orca Cetacea -Manatee/Dugong 1

Dugong Dugong dugon Sirenia

Manatee/Dugong 2

Caribbean Manatee Trichechus manatus Sirenia

Seal Harbor Seal Phoca vitulina CarnivoraSea Lion California Sea Lion Zalophus californianus CarnivoraWalrus Walrus Odobenus rosmarus CarnivoraSea Otter Sea Otter Enhydra lutris CarnivoraBear Polar Bear Ursus maritimus CarnivoraLand Carnivore 1 Eurasian Wolf Canis lupus lupus CarnivoraLand Carnivore 2 Dog Canis lupus familiaris CarnivoraArtiodactyl 1 Hippopotamus Hippopotamus amphibius ArtiodactylaArtiodactyl 2 Cow Bos taurus ArtiodactylaPerissodactyl 1 Horse Equus caballus Perissodacty

laPerissodactyl 2 Indian Rhinoceros Rhinoceros unicornis Perissodacty

laElephant 1 Asian Elephant Elephas maximus ProboscideaElephant 2 African Elephant Loxodonta africana Proboscidea

Fill in the first column entitled, “Relationship toWhales,” by assigning a number 1-13 for each species in terms of how closely you believe they are related to whales. (The species that you think is most closely related to whales gets a 1, the next closest relative gets a 2, the next a 3, etc.)

After modifying all of your sequences, be sure to save your Word Document!!!Part II – Sequence Alignments

13.A feature of the NCBI webpage is the Constraint Based Multiple Alignment Tool (COBALT). COBALT serves to use the resources of NCBI to compare sequences and put them into a progressive multiple alignment. From there, a simple phylogenic tree can be constructed from this alignment.

http://www.ncbi.nlm.nih.gov/tools/cobalt/

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14.The COBALT page asks you to enter protein ascensions, gis, or FASTA sequences. We have already compiled this information in the first part of this exercise. Return to your Word document that contains your cytochrome sequences. Select all of your sequences, Copy them and Paste them into the box where it indicates you should paste sequences.

15.The alignment itself takes several moments to run (more so today because we are all on the site at the same time). Once it has completed, it will show you the analysis that it ran between all species. The information provided will include information on things like number of amino acids in the protein and information about the similarities and differences that occur in the sequences. You can get the program to construct a phylogenetic tree by clicking on the “Phylogenetic Tree” hyperlink in the top right hand corner of the page.

A phylogenetic tree shows the evolutionary relationship among various species that are believed to have a common ancestor. These trees also take in evolutionary time into consideration through the distances between sequences (or the “leaves” of the tree).

16.In the space below, copy the phylogram you created in today’s class.

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Paste your sequences into this box.

Analysis:

How close were your predictions to your phylogram?

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Did you discover anything surprising about the relationship between whales and the other groups of mammals? If so, explain.

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The sequences you analyzed today were amino acid sequences. Would you expect to get the same results if you compared the DNA of these organisms instead of amino acids? Explain your response.

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In three or more sentences, use what you learned today to explain how protein similarity between species can be used to determine their evolutionary relationship.

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____________________________________________________________________________Sequences

>Blue.WhaleMTNIRKTHPLMKIINDAFIDLPTPSNISSWWNFGSLLGLCLIVQILTGLFLAMHYTPDTMTAFSSVTHIC

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RDVNYGWVIRYLHANGASMFFICLYAHMGRGLYYGSHAFRETWNIGVILLFTVMATAFVGYVLPWGQMSFWGATVITNLLSAIPYIGTTLVEWIWGGFSVDKATLTRFFAFHFILPFIIMALAIVHLIFLHETGSNNPTGIPSDMDKIPFHPYYTIKDILGALLLILTLLMLTLFAPDLLGDPDNYTPANPLSTPAHIKPEWYFLFAYAILRSIPNKLGGVLALLLSILVLALIPMLHTSKQRSMMFRPFSQFLFWVLVADLLTLTWIGGQPVEHPYVIVGQLASILYFLLILVLMPVTSLIENKLMKW

>Humpback.WhaleMTNIRKTHPLMKIINDTFIDLPTPSNISSWWNFGSLLGLCLIMQILTGLFLAMHYTPDTTTAFSSVTHICRDVNYGWIIRYLHANGASMFFICLYAHMGRGLYYGSYAFRETWNIGVILLFTVMATAFVGYVLPWGQMSFWGATVITNLLSAIPYIGTTLVEWIWGGFSVDKATLTRFFAFHFILPFIITALAIVHLIFLHETGSNNPTGIPSNMDKIPFHPYYTIKDTLGALLLILTLLMLTLFAPDLLGDPDNYTPANPLSTPAHIKPEWYFLFAYAILRSIPNKLGGVLALLLSILILAFIPMLHTSKQRSMMFRPFSQFLFWMLVADLLTLTWIGGQPVEHPYMIVGQLASILYFLLILVLMPMTSLIENKLMKW

>Sperm.WhaleMTNIRKSHPLMKIINNAFIDLPTPSNISSWWNFGSLLGLCLIMQILTGLFLAMHYTPDTTTAFSSITHICRDVNYGWTIRYLHANGASMFFICLYTHMGRGWYYGSYIFQETWNVGMMLLITVMATAFVGYVLPWGQMSFWAATVITNLLSAIPYIGTTLVEWVWGGFSVDKATLTRFFTLHFILPFITLTLTMVHLLFLHETGSNNPTGIPSNMDKIPFHPYHTIKDTMGALLLILSLLTLTLFAPDLLGDPDNYTPANPLNTPTHIKPEWYFLFAYAILRSVPNKLGGVLALLLSILILVFIPMLHTAKQRSMMFRPFSQFLFWTLIMDLLTLTWIGGQPVEHPYVTVGQLASILYFLLILILMPTASLIENKLLKW

>Killer.WhaleMTNIRKTHPLMKILNNAFIDLPTPSNISSWWNFGSLLGLCLITQILTGLLLAMHYTPDTSTAFSSVAHICRDVNYGWFIRYLHANGASMFFICLYAHIGRSLYYGSYMFQETWNVGVLLLLAVMATAFVGYVLPWGQMSFWGATVITNLLSAIPYIGTTLVEWIWGGFSVDKATLTRFFAFHFILPFIITALAAVHLLFLHETGSNNPTGIPSNMDMIPFHPYHTIKDTLGALLLILTLLALTLFAPDLLGDPDNYTPANPLSTPAHIKPEWYFLFAYAILRSVPNKLGGVLALLLSILILIFIPMLQTSKQRSMMFRPFSQLLFWTLIADLLTLTWIGGQPVEHPYIIVGQLASILYFLLILVLMPTISLIENKLLKW

>DugongMTNIRKSHPLIKILNNSFIDLPTPVNISSWWNFGSLLGACLIIQILTGLFLAMHYTSDTLTAFSSVTHICRDVNYGWIIRYLHANGASMFFLCLYAHIGRGIYYGSYLYPETWNIGIVLLLTVMATAFMGYVLPWGQMSFWGATVITNLLSAIPYIGTNLVEWVWGGFSVDKATLTRFFALHFILPFIVTALVMVHLLFLHETGSNNPTGLISDSDKIPFHPYYSVKDLLGLFLLILVLLLLTLFSPDMLGDPDNYTPANPLNTPPHIKPEWYFLFRYAILRSIPNKLGGVLALVLSILILALLPLLHTSKQRSLSFRPLSQCLFWILVADLITLTWIGGQPVEHPYIIIGQLASILYFSIILIFMPIAGLIENHLLKW

>Caribbean.Manatee MTNIRKSHPLIKILNDSFIDLPTPTNISSWWNFGSLLGACLIIQILTGLFLAMHYTSDTLTAFSSVAHICRDVNYGWIIRYLHANGASMFFLCLYAHIGRGIYYGSYLYSETWNIGIILLLTIMATAFMGYVLPWGQMSFWGATVITNLLSAIPYIGTNLVEWVWGGFSVDKATLTRFFALHFTLPFVVTALVTVHLLFLHETGSNNPTGLISDSDKIPFHPYYSIKDLLGLFLLILILLLPTLFSPDMLGDPDNYTPANPLSTPPHIKPEWYFLFAYAILRSIPNKLGGVLALVLSILILALLPLLHTSKQRSLLFRPLSQCLFWILVTDLITLTWIGGQPVEHPYIVIGQLASILYFSIILIFMPIAGLIENHLLKW

>Harbor.SealMTNIRKTHPLMKIINNSFIDLPTPSNISAWWNFGSLLGICLILQILTGLFLAMHYTSDTTTAFSSVTHICRDVNYGWIIRYLHANGASMFFICLYMHVGRGLYYGSYTFTETWNIGIILLFTVMATAFMGYVLPWGQMSFWGATVITNLLSAIPYVGTDLVQWIWGGFSVDKATLTRFFAFHFILPFVVLALDAVHLLFLHETGSNNPSGIMSDSDKIPFHPYYTIKDILGALLLILVLTLLVLFSPDLLGDPDNYIPPNPLSTPPHIKPEWYFLFAYAILRSIPNKLGGVLALVLSILVLAIMPLLHTSKQRGMMFRPISQCLFWFLVADLLTLTWIGGQPVEHPYITVGQLASILYFTILLVLMPIASIIENNILKW

>California.Sea.LionMTNIRKVHPLAKIINSSLIDLPTPSNISAWWNFGSLLAACLALQILTGLFLAMHYTSDTTTAFSSVTHICRDVNYGWIIRYMHANGASMFFICLYMHVGRGLYYGSYTLTETWNIGIILLFTIMATAFMGYVLPWGQMSFWGATVITNLLSAVPYIGTNLVEWIWGGFSVDKATLTRFFAFHFILPFMASALVMVHLLFLHETGSNNPSGISSDSDKIPFHPYYTIKDILGTLLLILTLMLLVMFSPDLLGDPDNYIPANPLSTPPHIKPEWYFLFAYAILRSIPNKLGGVLALLLSILILAIIPLLHTSKQRGMMFRPISQCLFWLLVADLLTLTWIGGQPVEHPFITIGQLASILYFTILLVFMPIAGIIENNILKW

>WalrusMTNIRKTHPLAKIINNTFIDLPTPSNISAWWNFGSLLATCLILQILTGLFLAMHYTSDTTTAFSSITHICRDVNYGWIIRYMHANGASMFFICLYAHMGRGIYYGSYTLAETWNIGIVLLLTIMATAFMGYVLPWGQMSFWGATVITNLLSAIPYVGTDLVEWVWGGFSVDKATLTRFLALHFVLPFMALALTAVHLLFLHETGSNNPSGILSDSDKIPFHPYYTIKDILGLIILILILMLLVLFSPDLLGDPDNYTPANPLSTPPHIKPEWYFLFAYAILRSIPNKLGGVLALLLSILVLAIVPSLHTSKQRSMMFRPISQCLFWLLVADLITLTWIGGQPVEHPFIIIGQLASILYFMILLVFMPIAGMIENSILKW

>Sea.OtterMTNIRKTHPLTKIINNSFIDLPAPSNISAWWNFGSLLGICLILQILTGLFLAMHYTSDTTTAFSSVAHICRDVNYGWIIRYMHANGASMFFICLFLHVGRGLYYGSYMFSETWNIGIVLLFTVMATAFMGYVLPWGQMSFWGATVITNLLSAIPYIGTNLVEWIWGGFSVDKATLTRFFAFHFILPFIISALAMIHLLFLHETGSNNPSGIPSNSDKIPFHPYYTIKDILGALSLVLALTTLVLFSPDLLGDPDNYIPANPLSTPPHIKPEWYFLFAYAILRSIPNKLGGVLALILSILILAIIPLLHTSKQRSMMFRPLSQCLFWLLVADLLTLTWIGGQPVEHPFVVIGQLASILYFMILLVLMPITSIIENNLLKW

>Polar.Bear

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MTNIRKTHPLAKIINNSFIDLPTPSNISAWWNFGSLLGVCLILQILTGLFLAMHYTSDTTTAFSSVTHICRDVHYGWVIRYVHANGASMFFICLFMHVGRGLYYGSYLFSETWNIGIILLFTVMATAFMGYVLPWGQMSFWGATVITNLLSAIPYIGTDLVEWIWGGFSVDKATLTRFFAFHFILPFIILALAAVHLLFLHETGSNNPSGIPSDSDKIPFHPYYTIKDILGALLLTLALATLVLFSPDLLGDPDNYIPANPLSTPPHIKPEWYFLFAYAILRSIPNKLGGVLALIFSILILALIPLLHTSKQRGMMFRPLSQCLFWLLVADLLTLTWIGGQPVEHPFIIIGQLASILYFTILLVLMPIAGIIENNLLKW

>Eurasian.WolfMTNIRKTHPLAKIVNNSFIDLPAPSNISAWWNFGSLLGVCLILQILTGLFLAMHYTSDTATAFSSVTHICRDVNYGWIIRYMHANGASMFFICLFLHVGRGLYYGSYVFMETWNIGIVLLFATMATAFMGYVLPWGQMSFWGATVITNLLSAIPYIGTDLVEWIWGGFSVDKATLTRFFAFHFILPFIIAALAMVHLLFLHETGSNNPSGITSDSDKIPFHPYYTIKDILGALLLLLILMSLVLFSPDLLGDPDNYTPANPLNTPPHIKPEWYFLFAYAILRSIPNKLGGVLALVFSILILAFIPLLHTSKQRSMMFRPLSQCLFWLLVADLLTLTWIGGQPVEHPFIIIGQVASMLYFTILLILMPTVSVIENNLLKW

>DogMTNIRKTHPLAKIVNNSFIDLPAPSNISAWWNFGSLLGVCLILQILTGLFLAMHYTSDTATAFSSVTHICRDVNYGWIIRYMHANGASMFFICLFLHVGRGLYYGSYVFMETWNIGIVLLFATMATAFMGYVLPWGQMSFWGATVITNLLSAIPYIGTDLVEWIWGGFSVDKATLTRFFAFHFILPFIIAALAMVHLLFLHETGSNNPSGITSDSDKIPFHPYYTIKDILGALLLLLILMSLVLFSPDLLGDPDNYTPANPLNTPPHIKPEWYFLFAYAILRSIPNKLGGVLALVFSILILAFIPLLHTSKQRSMMFRPLSQCLFWLLVADLLTLTWIGGQPVEHPFIIIGQVASILYFTILLILMPTVSVIENNLLKW

>HippopotamusMTNIRKSHPLMKIINDAFVDLPAPSNISSWWNFGSLLGVCLILQILTGLFLAMHYTPDTLTAFSSVTHICRDVNYGWVIRYMHANGASIFFICLFTHVGRGLYYGSHTFLETWNIGVILLLTTMATAFMGYVLPWGQMSFWGATVITNLLSAIPYIGTDLVEWIWGGFSVDKATLTRFFAFHFILPFVITALAIVHLLFLHETGSNNPTGIPSNADKIPFHPYYTIKDILGILLLMTTLLTLTLFAPDLLGDPDNYTPANPLSTPPHIKPEWYFLFAYAILRSIPNKLGGVLALALSILILALIPMLHTSKQRSLMFRPLSQCLFWALIADLLTLTWIGGQPVEHPFIIIGQVASILYFLLILVLMPVAGIIENKLLKW

>CowMTNIRKSHPLMKIVNNAFIDLPAPSNISSWWNFGSLLGICLILQILTGLFLAMHYTSDTTTAFSSVTHICRDVNYGWIIRYMHANGASMFFICLYMHVGRGLYYGSYTFLETWNIGVILLLTVMATAFMGYVLPWGQMSFWGATVITNLLSAIPYIGTNLVEWIWGGFSVDKATLTRFFAFHFILPFIIMAIAMVHLLFLHETGSNNPTGISSDVDKIPFHPYYTIKDILGALLLILALMLLVLFAPDLLGDPDNYTPANPLNTPPHIKPEWYFLFAYAILRSIPNKLGGVLALAFSILILALIPLLHTSKQRSMMFRPLSQCLFWALVADLLTLTWIGGQPVEHPYITIGQLASILYFLLILVLMPTAGTIENKLLKW

>HorseMTNIRKSHPLIKIINHSFIDLPAPSNISSWWNFGSLLGICLILQILTGLFLAMHYTSDTTTAFSSVTHICRDVNYGWIIRYLHANGASMFFICLFIHVGRGLYYGSYTFLETWNIGIILLFTVMATAFMGYVLPWGQMSFWGATVITNLLSAIPYIGTTLVEWIWGGFSVDKATLTRFFAFHFILPFIITALVVVHLLFLHETGSNNPSGIPSDMDKIPFHPYYTIKDILGLLLLILLLLTLVLFSPDLLGDPDNYTPANPLSTPPHIKPEWYFLFAYAILRSIPNKLGGVLALILSILILALIPTLHMSKQRSMMFRPLSQCVFWLLVADLLTLTWIGGQPVEHPYVIIGQLASILYFSLILIFMPLASTIENNLLKW

>RhinocerosMTNIRKSHPLVKIINHSFIDLPTPSNISSWWNFGSLLGICLILQILTGLFLAMHYTPDTTTAFSSVTHICRDVNYGWMIRYLHANGASMFFICLFIHVGRGLYYGSYTFLETWNIGIILLFTLMATAFMGYVLPWGQMSFWGATVITNLLSAIPYIGTNLVEWIWGGFSVDKATLTRFFAFHFILPFIILALAITHLLFLHETGSNNPSGIPSNMDKIPFHPYYTIKDILGALLLILVLLILVLFFPDILGDPDNYTPANPLSTPPHIKPEWYFLFAYAILRSIPNKLGGVLALAFSILILLLIPYLHTSKQRSMMFRPLSQCMFWLLVADLLTLTWIGGQPVEHPFIIIGQLASILYFSLILVLMPLAGIIENNLLKW

>Indian.ElephantMTHTRKSHPLFKIINKSFIDLPTPSNISTWWNFGSLLGACLITQILTGLFLAMHYTPDTMTAFSSMSHICRDVNYGWIIRQLHSNGASIFFLCLYTHIGRNIYYGSYLYSETWNTGIMLLLITMATAFMGYVLPWGQMSFWGATVITNLFSAIPYIGTNLVEWIWGGFSVDKATLNRFFAFHFILPFTMVALAGVHLTFLHETGSNNPLGLTSNSDKIPFHPYYTIKDFLGLLILILLLLLLALLSPDMLGDPDNYMPADPLNTPLHIKPEWYFLFAYAILRSVPNKLGGVLALFLSILILGLMPFLHTSKHRSMMLRPLSQVLFWTLTMDLLTLTWIGSQPVEYPYTIIGQMASILYFSIILAFLPIAGMIENYLIK

>African.ElephantMTHIRKSHPLLKIINKSFIDLPTPSNISTWWNFGSLLGACLITQILTGLFLAMHYTPDTMTAFSSMSHICRDVNYGWIIRQLHSNGASIFFLCLYTHIGRNIYYGSYLYSETWNTGIMLLLITMATAFMGYVLPWGQMSFWGATVITNLFSAIPYIGTNLVEWIWGGFSVDKATLNRFFALHFILPFTMIALAGVHLTFLHETGSNNPLGLTSDSDKIPFHPYYTIKDFLGLLILILLLLLLALLSPDMLGDPDNYMPADPLNTPLHIKPEWYFLFAYAILRSVPNKLGGVLALLLSILILGLMPLLHTSKHRSMMLRPLSQVLFWTLTMDLLTLTWIGSQPVEYPYIIIGQMASILYFSIILAFLPIAGVIENYLIK

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