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Using the protein sequence for human beta globin do a BLASTP search in the UCSD Biology WorkBench using the SwissProt database. Using CLUSTALW and other tools we learned in this weeks tutorials, answer the following: Based on the CLUSTALW alignment and DRAWTREE evolutionary trees generated from the alignment of beta globin proteins from mountain gorilla, pigs, rabbits, dogs, badger, otter, bats, pandas and chimpanzees, what conclusions can you draw about the evolution of beta globin? Can you use this information to extrapolate about the evolutionary histories of these animal lineages? EXPLAIN why or why not? (click here for organism ID information) I was unable to find the required species from the available listings (GORBE, CANFA, ROUAE did not appear anywhere in the list of returned options), therefore, I selected the following: Selected Sequence(s) Hemoglobin subunit beta-1/2 [Oryctolagus cuniculus (Rabbit)] Hemoglobin subunit beta [Ailuropoda melanoleuca (Giant panda)] Hemoglobin subunit beta [Meles meles (Eurasian badger)] Hemoglobin subunit beta [Lutra lutra (European river otter)] Hemoglobin subunit epsilon [Sus scrofa (Pig)] Hemoglobin subunit epsilon [Gorilla gorilla gorilla (Lowland gorilla)] Hemoglobin subunit epsilon [Pan troglodytes (Chimpanzee)] Hemoglobin subunit epsilon [Macaca mulatta (Rhesus macaque)] Hemoglobin subunit epsilon [Pongo pygmaeus (Bornean orangutan)] Consensus key (see documentation for details) * - single, fully conserved residue : - conservation of strong groups . - conservation of weak groups - no consensus CLUSTAL W (1.81) multiple sequence alignment HBE_PANTR HBE_GORGO HBE_PONPY HBE_MACMU HBE_PIG HBB_LUTLU HBB_AILME HBB_MELME HBB_RABIT MVHFTAEEKAAVTSLWSKMNVEEAGGEALGRLLVVYPWTQRFFDSFGNLSSPSAILGNPK MVHFTAEEKAAVTSLWSKMNVEEAGGEALGRLLVVYPWTQRFFDSFGNLSSPSAILGNPK MVHFTAEEKAAVTSLWSKMNVEEAGGEALGRLLVVYPWTQRFFDSFGNLSSPSAILGNPK MVHFTAEEKAAVTSLWSKMNVEETGGEALGRLLVVYPWTQRFFDSFGNLSSPSAILGNPK MVHFTAEEKSVITGLWGKVNVEETGGQAVGRLLVVYPWTQRFFDSFGNMSSPSAIMGNPK -VHLTGEEKAAVTSLWGKVNVDEVGGEALGRLLVVYPWTQRFFDSFGDLSSPDAVMGNPK MVHLTGEEKAAVTGLWSKVNVDEVGGEALGRLLVVYPWTQRFFDSFGDLSTPDAVMNNPK -VHLTAEEKSAVTSLWGKVNVDEVGGEALGRLLVVYPWTQRYFDSFGDLSTPDAVMGNPK MVHLSSEEKSAVTALWGKVNVEEVGGEALGRLLVVYPWTQRFFESFGDLSSANAVMNNPK **::.***:.:*.**.*:**:*.**:*:************:*:***::*:..*::.*** HBE_PANTR HBE_GORGO HBE_PONPY HBE_MACMU HBE_PIG HBB_LUTLU HBB_AILME VKAHGKKVLTSFGDAIKNMDNLKPAFAKLSELHCDKLHVDPENFKLLGNVMVIILATHFG VKAHGKKVLTSFGDAIKNMDNLKPAFAKLSELHCDKLHVDPENFKLLGNVMVIILATHFG VKAHGKKVLTSFGDAIKNMDNLKTTFAKLSELHCDKLHVDPENFKLLGNVMVIILATHFG VKAHGKKVLTSFGDAIKNMDNLKITFAKLSELHCDKLHVDPENFKLLGNVMVIILATHFG VKAHGKKVLTAFGDAVKNMDNLKGTFAKLSELHCDKLHVDPENFRLLGNMIVIILASHFG VKAHGKKVLNSFSEGLKNLDNLKGTFAKLSELHCDKLHVDPENFKLLGNVLVCVLAHHFG VKAHGKKVLNSFSEGLKNLDNLKGTFAKLSELHCDKLHVDPENFKLLGNVLVCVLAHHFG HBB_MELME HBB_RABIT VKAHGKKVLNSFSEGLKNLDNLKGTFAKLSELHCDKLHVDPENFKLLGNVLVCVLAHHFG VKAHGKKVLAAFSEGLSHLDNLKGTFAKLSELHCDKLHVDPENFRLLGNVLVIVLSHHFG ********* :*.:.:.::**** :*******************:****::* :*: *** HBE_PANTR HBE_GORGO HBE_PONPY HBE_MACMU HBE_PIG HBB_LUTLU HBB_AILME HBB_MELME HBB_RABIT KEFTPEVQAAWQKLVSAVAIALAHKYH KEFTPEVQAAWQKLVSAVAIALAHKYH KEFTPEVQAAWQKLVSAVAIALAHKYH KEFTPEVQAAWQKLVSAVAIALAHKYH GEFTPEVQAAWQKLVAGVATALAHKYH KEFTPQVQAAYQKVVAGVANALAHKYH KEFTPQVQAAYQKVVAGVANALAHKYH KEFTPQVQAAYQKVVAGVANALAHKYH KEFTPQVQAAYQKVVAGVANALAHKYH ****:****:**:*:.** ******* a. Paste your BOXSHADE output here: b. Paste your unrooted tree here: c. Paste your rooted tree here: Given the above data, I’d propose that the evolutionary histories of these animal lineages are quite similar, as far as the evolution of beta-globin is concerned. As indicated by the BOXSHADE output, the alignment of the beta-globin amino acid sequence is very often fully conserved across each species. Both the unrooted and rooted phylogenetic trees show greater segmentation among the selected species. PANTR, GORGO, PONPY and MACMU are all relatively close in their genetic makeup of beta-globin, whereas LUTLU, AILME, MELME and RABIT share closeness with each other in a cluster separate from the primate group. PIG represents a section of species equidistant between the ruminants and primates, to the extent of evolutionary divergence of the beta-globin gene. 2. Go back and BLAST the myoglobin sequence as in the tutorial. Produce a rooted and unrooted tree for vertebrates from a multiple alignment pick the same species as above. I recognize that the same species have not been selected, however, as there were different workbench labels when aligning the different proteins, I had to select what was available. Selected Sequence(s) Myoglobin [Ailuropoda melanoleuca (Giant panda)] Myoglobin [Meles meles (Eurasian badger)] Myoglobin [Oryctolagus cuniculus (Rabbit)] Myoglobin [Lutra lutra (European river otter)] Myoglobin [Sus scrofa (Pig)] Myoglobin [Macaca fascicularis (Crab-eating macaque) (Cynomolgus monkey)] Myoglobin [Pongo pygmaeus (Bornean orangutan)] Myoglobin [Gorilla gorilla beringei (Mountain gorilla) (Highland gorilla)] Myoglobin [Pan troglodytes (Chimpanzee)] Consensus key (see documentation for details) * - single, fully conserved residue : - conservation of strong groups . - conservation of weak groups - no consensus CLUSTAL W (1.81) multiple sequence alignment MYG_PANTR MYG_GORBE MYG_PONPY MYG_MACFA MYG_PIG MYG_RABIT MYG_MELME MYG_AILME MYG_LUTLU MYG_PANTR MYG_GORBE MYG_PONPY MYG_MACFA MYG_PIG MYG_RABIT MYG_MELME MYG_AILME MYG_LUTLU MYG_PANTR MYG_GORBE MYG_PONPY MYG_MACFA MYG_PIG MYG_RABIT MGLSDGEWQLVLNVWGKVEADIPGHGQEVLIRLFKGHPETLEKFDKFKHLKSEDEMKASE MGLSDGEWQLVLNVWGKVEADISGHGQEVLIRLFKGHPETLEKFDKFKHLKSEDEMKASE MGLSDGEWQLVLNVWGKVEADIPSHGQEVLIRLFKGHPETLEKFDKFKHLKSEDEMKASE MGLSDGEWQLVLNVWGKVEADIPSHGQEVLIRLFKGHPETLEKFDKFKHLKSEDEMKASE MGLSDGEWQLVLNVWGKVEADVAGHGQEVLIRLFKGHPETLEKFDKFKHLKSEDEMKASE MGLSDAEWQLVLNVWGKVEADLAGHGQEVLIRLFHTHPETLEKFDKFKHLKSEDEMKASE MGLSDGEWQLVLNVWGKVEADLAGHGQEVLIRLFKGHPETLEKFDKFKHLKSEDEMKGSE -GLSDGEWQLVLNVWGKVEADLAGHGQEVLIRLFKGHPETLEKFDKFKHLKSE---KGSE MGLSDGEWQLVLNVWGKVEADLAGHGQEVLIRLFKGHPETLEKFDKFKHLKSEDEMKGSE ****.***************:..**********: ***************** *.** DLKKHGATVLTALGGILKKKGHHEAEIKPLAQSHATKHKIPVKYLEFISECIIQVLHSKH DLKKHGATVLTALGGILKKKGHHEAEIKPLAQSHATKHKIPVKYLEFISECIIQVLQSKH DLKKHGATVLTALGGILKKKGHHEAEIKPLAQSHATKHKIPVKYLEFISESIIQVLQSKH DLKKHGVTVLTALGGILKKKGHHEAEIKPLAQSHATKHKIPVKYLELISESIIQVLQSKH DLKKHGNTVLTALGGILKKKGHHEAELTPLAQSHATKHKIPVKYLEFISEAIIQVLQSKH DLKKHGNTVLTALGAILKKKGHHEAEIKPLAQSHATKHKIPVKYLEFISEAIIHVLHSKH DLKKHGNTVLTALGGILKKKGHQEAELKPLAQSHATKHKIPVKYLEFISDAIAQVLQSKH DLKKHGNTVETALEGILKKK---------------------------------------DLKKHGNTVLTALGGILKKKGKHEAELKPLAQSHATKHKIPIKYLEFISEAIIQVLQSKH ****** ** *** .***** PGDFGADAQGAMNKALELFRKDMASNYKELGFQG PGDFGADAQGAMNKALELFRKDMASNYKELGFQG PGDFGADAQGAMNKALELFRKDMASNYKELGFQG PGDFGADAQGAMNKALELFRNDMAAKYKELGFQG PGDFGADAQGAMSKALELFRNDMAAKYKELGFQG PGDFGADAQAAMSKALELFRNDIAAQYKELGFQG MYG_MELME MYG_AILME MYG_LUTLU PGNFAAEAQGAMKKALELFRNDIAAKYKELGFQG --------------ALELFKNDIAAKTKELGFLG PGDFGADAQGAMKRALELFRNDIAAKYKELGFQG *****::*:*:: ***** * a. Paste your BOXSHADE output here: b. Paste your unrooted tree here: c. Paste your rooted tree here: 3. Is the beta globin tree the same as the myoglobin tree? EXPLAIN what this means with regard to inferring evolutionary relationships among these species. Although the trees are similar, they are not the same. These trees each represent the phylogenesis of the different species, and as such, depict the ancestry of the species in regards to the development of their myologin and beta-globin genes. In other words, we cannot tell the proximity of relationship from the trees, but we can infer, for example, that MELME and AILME are more closely related in their genetics than RABIT and LUTLU.