Amino Acid Found in the Protein - Assignment Example

Name: Instructor: Course: Date: Sequence ATGGGATTTGAGATTGCAAAGACCAACTCAATCTTATCAAAATTGGCTACTAATGAAGAGCATGGCGAAAACTCGCCATATTTTGATGGGTGGAAAGCATACGATAGTGATCCTTTCCACCCTCTAAAAAACCCCAACGGAGTTATCCAAATGGGTCTTGCTGAAAATCAGCTTTGTTTAGACTTGATAGAAGATTGGATTAAGAGAAACCCAAAAGGTTCAATTTGTTCTGAAGGAATCAAATCATTCAAGGCCATTGCCAACTTTCAAGATTATCATGGCTTGCCTGAATTCAGAAAAGCGATTGCGAAATTTATGGAGAAAACAAGAGGAGGAAGAGTTAGATTTGATCCAGAAAGAGTTGTTATGGTTGGTGGTGCCACTGGAGCTAATGAGACAATTATATTTTGTTTGGCTGATCCTGGCGATGCATTTTTAGTACCTTCACCATACTACCCAGCATTTAACAGAGATTTAAGATGGAGAACTGGAGTACAACTTATTCCAATTCACTGTGAGAGCTCCAATAATTTCAAAATTACTTCAAAAGCAGTAAAAGAAGCATATGAAAATGCACAAAAATCAAACATCAAAGTAAAAGGTTTGATTTTGACCAATCCATCAAATCCATTGGGCACCACTTTGGACAAAGACACACTGAAAAGTGTCTTGAGTTTCACCAACCAACACAACATCCACCTTGTTTGTGACGAAATCTACGCAGCCACTGTCTTTGACACGCCTCAATTCGTCAGTATAGCTGAAATCCTCGATGAACAGGAAATGACTTACTGCAACAAAGATTTAGTTCACATCGTCTACAGTCTTTCAAAAGACATGGGGTTACCAGGATTTAGAGTCGGAATCATATATTCTTTTAACGACGATGTCGTTAATTGTGCTAGAAAAATGTCGAGTTTCGGTTTAGTATCTACACAAACGCAATATTTTTTAGCGGCAATGCTATCGGACGAAAAATTCGTCGATAATTTTCTAAGAGAAAGCGCGATGAGGTTAGGTAAAAGGCACAAACATTTTACTAATGGACTTGAAGTAGTGGGAATTAAATGCTTGAAAAATAATGCGGGGCTTTTTTGTTGGATGGATTTGCGTCCACTTTTAAGGGAATCGACTTTCGATAGCGAAATGTCGTTATGGAGAGTTATTATAAACGATGTTAAGCTTAACGTCTCGCCTGGATCTTCGTTTGAATGTCAAGAGCCAGGGTGGTTCCGAGTTTGTTTTGCAAATATGGATGATGGAACGGTTGATATTGCGCTCGCGAGGATTCGGAGGTTCGTAGGTGTTGAGAAAAGTGGAGATAAATCGAGTTCGATGGAAAAGAAGCAACAATGGAAGAAGAATAATTTGAGACTTAGTTTTTCGAAAAGAATGTATGATGAAAGTGTTTTGTCACCACTTTCGTCACCTATTCCTCCCTCACCATTAGTTCGTTAA >gi|19167|emb|X59145.1| Lycopersicon esculentum LE-ASCC2 mRNA (ptACC2) for 1-aminocyclopropane-1-carboxylic acid synthase TGTAGTTGTGTACATTTTATTAATCTTCATCTTCTTAATTCTCTTCAGTTTTTAATTTCTTCACTTCTAA ACTCATTTAGTAAAAAAAAAATGGGATTTGAGATTGCAAAGACCAACTCAATCTTATCAAAATTGGCTAC TAATGAAGAGCATGGCGAAAACTCGCCATATTTTGATGGGTGGAAAGCATACGATAGTGATCCTTTCCAC CCTCTAAAAAACCCCAACGGAGTTATCCAAATGGGTCTTGCTGAAAATCAGCTTTGTTTAGACTTGATAG AAGATTGGATTAAGAGAAACCCAAAAGGTTCAATTTGTTCTGAAGGAATCAAATCATTCAAGGCCATTGC CAACTTTCAAGATTATCATGGCTTGCCTGAATTCAGAAAAGCGATTGCGAAATTTATGGAGAAAACAAGA GGAGGAAGAGTTAGATTTGATCCAGAAAGAGTTGTTATGGTTGGTGGTGCCACTGGAGCTAATGAGACAA TTATATTTTGTTTGGCTGATCCTGGCGATGCATTTTTAGTACCTTCACCATACTACCCAGCATTTAACAG AGATTTAAGATGGAGAACTGGAGTACAACTTATTCCAATTCACTGTGAGAGCTCCAATAATTTCAAAATT ACTTCAAAAGCAGTAAAAGAAGCATATGAAAATGCACAAAAATCAAACATCAAAGTAAAAGGTTTGATTT TGACCAATCCATCAAATCCATTGGGCACCACTTTGGACAAAGACACACTGAAAAGTGTCTTGAGTTTCAC CAACCAACACAACATCCACCTTGTTTGTGACGAAATCTACGCAGCCACTGTCTTTGACACGCCTCAATTC GTCAGTATAGCTGAAATCCTCGATGAACAGGAAATGACTTACTGCAACAAAGATTTAGTTCACATCGTCT ACAGTCTTTCAAAAGACATGGGGTTACCAGGATTTAGAGTCGGAATCATATATTCTTTTAACGACGATGT CGTTAATTGTGCTAGAAAAATGTCGAGTTTCGGTTTAGTATCTACACAAACGCAATATTTTTTAGCGGCA ATGCTATCGGACGAAAAATTCGTCGATAATTTTCTAAGAGAAAGCGCGATGAGGTTAGGTAAAAGGCACA AACATTTTACTAATGGACTTGAAGTAGTGGGAATTAAATGCTTGAAAAATAATGCGGGGCTTTTTTGTTG GATGGATTTGCGTCCACTTTTAAGGGAATCGACTTTCGATAGCGAAATGTCGTTATGGAGAGTTATTATA AACGATGTTAAGCTTAACGTCTCGCCTGGATCTTCGTTTGAATGTCAAGAGCCAGGGTGGTTCCGAGTTT GTTTTGCAAATATGGATGATGGAACGGTTGATATTGCGCTCGCGAGGATTCGGAGGTTCGTAGGTGTTGA GAAAAGTGGAGATAAATCGAGTTCGATGGAAAAGAAGCAACAATGGAAGAAGAATAATTTGAGACTTAGT TTTTCGAAAAGAATGTATGATGAAAGTGTTTTGTCACCACTTTCGTCACCTATTCCTCCCTCACCATTAG TTCGTTAAGACTTAATTAAAAGGGAAGAATTTAATTTATGTTTTTTTATATTTTGAAAAAAATTTGTAAG AATAAGATTATAATAGGAAAAGAAAATAAGTATGTAGGATGAGGAGTATTTTCAGAAATAGTTGTTAGCG TATGTATTGACAACTGGTCTATGTACTTAGACATCATAATTTGTCTTAGCTAATTAACGAATGCAAAAGT GAAGTTATGTTATGACTCTTAGAAT What is the name of the gene that you identified by your GenBank search? The name of the gene is LE-ASCC2 mRNA (ptACC2) or 1-aminocyclopropane-1-carboxylatesynthase gene What organism is this gene from? This particular gene is from Solanum lycopersicum (Lycopersicon esculentum) What is the name of the protein encoded by this gene? Depending on your sequence, this may be the same name as the gene. The protein is called 1-aminocyclopropane-1-carboxylic acid synthase. What is the sequence of the first ten amino acids in the protein encoded by this open reading frame? 1-aminocyclopropane 1-carboxylate synthase [Solanum lycopersicum] GenBank: CAA41856.1 >gi|19168|emb|CAA41856.1| 1-aminocyclopropane 1-carboxylate synthase [Solanum lycopersicum] MGFEIAKTNSILSKLATNEEHGENSPYFDGWKAYDSDPFHPLKNPNGVIQMGLAENQLCLDLIEDWIKRN PKGSICSEGIKSFKAIANFQDYHGLPEFRKAIAKFMEKTRGGRVRFDPERVVMVGGATGANETIIFCLAD PGDAFLVPSPYYPAFNRDLRWRTGVQLIPIHCESSNNFKITSKAVKEAYENAQKSNIKVKGLILTNPSNP LGTTLDKDTLKSVLSFTNQHNIHLVCDEIYAATVFDTPQFVSIAEILDEQEMTYCNKDLVHIVYSLSKDM GLPGFRVGIIYSFNDDVVNCARKMSSFGLVSTQTQYFLAAMLSDEKFVDNFLRESAMRLGKRHKHFTNGL EVVGIKCLKNNAGLFCWMDLRPLLRESTFDSEMSLWRVIINDVKLNVSPGSSFECQEPGWFRVCFANMDD GTVDIALARIRRFVGVEKSGDKSSSMEKKQQWKKNNLRLSFSKRMYDESVLSPLSSPIPPSPLVR The sequence of the first ten amino acids in this protein as encoded in the open reading frame is: Methionine, Glycine, Phenylalanine, Glutamate, Isoleucine, Alanine, Lysine, Threonine, Asparagine, Serine, 1-aminocyclopropane-1-carboxylate synthase gene sequence: 1458 base pairs atgggatttgagattgcaaagaccaactcaatcttatcaaaattggctactaatgaagagcatggcgaaaactcg base pairs taccctaaactctaacgtttctggttgagttagaatagttttaaccgatgattacttctcgtaccgcttttgagc 1 to 75 ccatattttgatgggtggaaagcatacgatagtgatcctttccaccctctaaaaaaccccaacggagttatccaa base pairs ggtataaaactacccacctttcgtatgctatcactaggaaaggtgggagattttttggggttgcctcaataggtt 76 to 150 atgggtcttgctgaaaatcagctttgtttagacttgatagaagattggattaagagaaacccaaaaggttcaatt base pairs tacccagaacgacttttagtcgaaacaaatctgaactatcttctaacctaattctctttgggttttccaagttaa 151 to 225 EcoRI tgttctgaaggaatcaaatcattcaaggccattgccaactttcaagattatcatggcttgcctgaattcagaaaa base pairs acaagacttccttagtttagtaagttccggtaacggttgaaagttctaatagtaccgaacggacttaagtctttt 226 to 300 gcgattgcgaaatttatggagaaaacaagaggaggaagagttagatttgatccagaaagagttgttatggttggt base pairs cgctaacgctttaaatacctcttttgttctcctccttctcaatctaaactaggtctttctcaacaataccaacca 301 to 375 ggtgccactggagctaatgagacaattatattttgtttggctgatcctggcgatgcatttttagtaccttcacca base pairs ccacggtgacctcgattactctgttaatataaaacaaaccgactaggaccgctacgtaaaaatcatggaagtggt 376 to 450 tactacccagcatttaacagagatttaagatggagaactggagtacaacttattccaattcactgtgagagctcc base pairs atgatgggtcgtaaattgtctctaaattctacctcttgacctcatgttgaataaggttaagtgacactctcgagg 451 to 525 aataatttcaaaattacttcaaaagcagtaaaagaagcatatgaaaatgcacaaaaatcaaacatcaaagtaaaa base pairs ttattaaagttttaatgaagttttcgtcattttcttcgtatacttttacgtgtttttagtttgtagtttcatttt 526 to 600 ggtttgattttgaccaatccatcaaatccattgggcaccactttggacaaagacacactgaaaagtgtcttgagt base pairs ccaaactaaaactggttaggtagtttaggtaacccgtggtgaaacctgtttctgtgtgacttttcacagaactca 601 to 675 ttcaccaaccaacacaacatccaccttgtttgtgacgaaatctacgcagccactgtctttgacacgcctcaattc base pairs aagtggttggttgtgttgtaggtggaacaaacactgctttagatgcgtcggtgacagaaactgtgcggagttaag 676 to 750 gtcagtatagctgaaatcctcgatgaacaggaaatgacttactgcaacaaagatttagttcacatcgtctacagt base pairs cagtcatatcgactttaggagctacttgtcctttactgaatgacgttgtttctaaatcaagtgtagcagatgtca 751 to 825 ctttcaaaagacatggggttaccaggatttagagtcggaatcatatattcttttaacgacgatgtcgttaattgt base pairs gaaagttttctgtaccccaatggtcctaaatctcagccttagtatataagaaaattgctgctacagcaattaaca 826 to 900 gctagaaaaatgtcgagtttcggtttagtatctacacaaacgcaatattttttagcggcaatgctatcggacgaa base pairs cgatctttttacagctcaaagccaaatcatagatgtgtttgcgttataaaaaatcgccgttacgatagcctgctt 901 to 975 aaattcgtcgataattttctaagagaaagcgcgatgaggttaggtaaaaggcacaaacattttactaatggactt base pairs tttaagcagctattaaaagattctctttcgcgctactccaatccattttccgtgtttgtaaaatgattacctgaa 976 to 1050 gaagtagtgggaattaaatgcttgaaaaataatgcggggcttttttgttggatggatttgcgtccacttttaagg base pairs cttcatcacccttaatttacgaactttttattacgccccgaaaaaacaacctacctaaacgcaggtgaaaattcc 1051 to 1125 HindIII gaatcgactttcgatagcgaaatgtcgttatggagagttattataaacgatgttaagcttaacgtctcgcctgga base pairs cttagctgaaagctatcgctttacagcaatacctctcaataatatttgctacaattcgaattgcagagcggacct 1126 to 1200 tcttcgtttgaatgtcaagagccagggtggttccgagtttgttttgcaaatatggatgatggaacggttgatatt base pairs agaagcaaacttacagttctcggtcccaccaaggctcaaacaaaacgtttatacctactaccttgccaactataa 1201 to 1275 gcgctcgcgaggattcggaggttcgtaggtgttgagaaaagtggagataaatcgagttcgatggaaaagaagcaa base pairs cgcgagcgctcctaagcctccaagcatccacaactcttttcacctctatttagctcaagctaccttttcttcgtt 1276 to 1350 caatggaagaagaataatttgagacttagtttttcgaaaagaatgtatgatgaaagtgttttgtcaccactttcg base pairs gttaccttcttcttattaaactctgaatcaaaaagcttttcttacatactactttcacaaaacagtggtgaaagc 1351 to 1425 tcacctattcctccctcaccattagttcgttaa base pairs agtggataaggagggagtggtaatcaagcaatt 1426 to 1458 Table by Enzyme Name Enzyme No. Positions Recognition name cuts of sites sequence EcoRI 1 289 g/aattc HindIII 1 1180 a/agctt The following endonucleases were selected but don't cut this sequence: BamHI How many BamHI sites are in the gene sequence? List the positions of any BamHI sites in the gene. BamHI sites are not in the sequence. How many EcoRI sites are in the gene sequence? List the positions of any EcoRI sites in the gene. EcoRI has one cutting site in the gene sequence at position 289. How many HindIII sites are in the gene sequence? List the positions of any HindIII sites in the gene. HindIII has one cutting site at position 1180 on the gene sequence. Mutated gene ATGGGATTTGAGATTGCAAAGACCAACTCAATCTTATCAAAATTGGCTACTAATGAAGAGCATGGCGAAAACTCGCCATATTTTGATGGGTGGAAAGCATACGATAGGGATCCTTTCCACCCTCTAAAAAACCCCAACGGAGTTATCCAAATGGGTCTTGCTGAAAATCAGCTTTGTTTAGACTTGATAGAAGATTGGATTAAGAGAAACCCAAAAGGTTCAATTTGTTCTGAAGGAATCAAATCATTCAAGGCCATTGCCAACTTTCAAGATTATCATGGCTTGCCTGAATTCAGAAAAGCGATTGCGAAATTTATGGAGAAAACAAGAGGAGGAAGAGTTAGATTTGATCCAGAAAGAGTTGTTATGGTTGGTGGTGCCACTGGAGCTAATGAGACAATTATATTTTGTTTGGCTGATCCTGGCGATGCATTTTTAGTACCTTCACCATACTACCCAGCATTTAACAGAGATTTAAGATGGAGAACTGGAGTACAACTTATTCCAATTCACTGTGAGAGCTCCAATAATTTCAAAATTACTTCAAAAGCAGTAAAAGAAGCATATGAAAATGCACAAAAATCAAACATCAAAGTAAAAGGTTTGATTTTGACCAATCCATCAAATCCATTGGGCACCACTTTGGACAAAGACACACTGAAAAGTGTCTTGAGTTTCACCAACCAACACAACATCCACCTTGTTTGTGACGAAATCTACGCAGCCACTGTCTTTGACACGCCTCAATTCGTCAGTATAGCTGAAATCCTCGATGAACAGGAAATGACTTACTGCAACAAAGATTTAGTTCACATCGTCTACAGTCTTTCAAAAGACATGGGGTTACCAGGATTTAGAGTCGGAATCATATATTCTTTTAACGACGATGTCGTTAATTGTGCTAGAAAAATGTCGAGTTTCGGTTTAGTATCTACACAAACGCAATATTTTTTAGCGGCAATGCTATCGGACGAAAAATTCGTCGATAATTTTCTAAGAGAAAGCGCGATGAGGTTAGGTAAAAGGCACAAACATTTTACTAATGGACTTGAAGTAGTGGGAATTAAATGCTTGAAAAATAATGCGGGGCTTTTTTGTTGGATGGATTTGCGTCCACTTTTAAGGGAATCGACTTTCGATAGCGAAATGTCGTTATGGAGAGTTATTATAAACGATGTTAAGCTTAACGTCTCGCCTGGATCTTCGTTTGAATGTCAAGAGCCAGGGTGGTTCCGAGTTTGTTTTGCAAATATGGATGATGGAACGGTTGATATTGCGCTCGCGAGGATTCGGAGGTTCGTAGGTGTTGAGAAAAGTGGAGATAAATCGAGTTCGATGGAAAAGAAGCAACAATGGAAGAAGAATAATTTGAGACTTAGTTTTTCGAAAAGAATGTATGATGAAAGTGTTTTGTCACCACTTTCGTCACCTATTCCTCCCTCACCATTAGTTCGTTAA What is the nucleotide base immediately 5' to the GATCC subsequence you detected, and at what position is it (using the first nucleotide in your sequence as 1)? The nucleotide base immediately upsteam on the 5’of GATCC is T Mutated gene sequence: 1458 base pairs atgggatttgagattgcaaagaccaactcaatcttatcaaaattggctactaatgaagagcatggcgaaaactcg base pairs taccctaaactctaacgtttctggttgagttagaatagttttaaccgatgattacttctcgtaccgcttttgagc 1 to 75 BamHI ccatattttgatgggtggaaagcatacgatagggatcctttccaccctctaaaaaaccccaacggagttatccaa base pairs ggtataaaactacccacctttcgtatgctatccctaggaaaggtgggagattttttggggttgcctcaataggtt 76 to 150 atgggtcttgctgaaaatcagctttgtttagacttgatagaagattggattaagagaaacccaaaaggttcaatt base pairs tacccagaacgacttttagtcgaaacaaatctgaactatcttctaacctaattctctttgggttttccaagttaa 151 to 225 EcoRI tgttctgaaggaatcaaatcattcaaggccattgccaactttcaagattatcatggcttgcctgaattcagaaaa base pairs acaagacttccttagtttagtaagttccggtaacggttgaaagttctaatagtaccgaacggacttaagtctttt 226 to 300 gcgattgcgaaatttatggagaaaacaagaggaggaagagttagatttgatccagaaagagttgttatggttggt base pairs cgctaacgctttaaatacctcttttgttctcctccttctcaatctaaactaggtctttctcaacaataccaacca 301 to 375 ggtgccactggagctaatgagacaattatattttgtttggctgatcctggcgatgcatttttagtaccttcacca base pairs ccacggtgacctcgattactctgttaatataaaacaaaccgactaggaccgctacgtaaaaatcatggaagtggt 376 to 450 tactacccagcatttaacagagatttaagatggagaactggagtacaacttattccaattcactgtgagagctcc base pairs atgatgggtcgtaaattgtctctaaattctacctcttgacctcatgttgaataaggttaagtgacactctcgagg 451 to 525 aataatttcaaaattacttcaaaagcagtaaaagaagcatatgaaaatgcacaaaaatcaaacatcaaagtaaaa base pairs ttattaaagttttaatgaagttttcgtcattttcttcgtatacttttacgtgtttttagtttgtagtttcatttt 526 to 600 ggtttgattttgaccaatccatcaaatccattgggcaccactttggacaaagacacactgaaaagtgtcttgagt base pairs ccaaactaaaactggttaggtagtttaggtaacccgtggtgaaacctgtttctgtgtgacttttcacagaactca 601 to 675 ttcaccaaccaacacaacatccaccttgtttgtgacgaaatctacgcagccactgtctttgacacgcctcaattc base pairs aagtggttggttgtgttgtaggtggaacaaacactgctttagatgcgtcggtgacagaaactgtgcggagttaag 676 to 750 gtcagtatagctgaaatcctcgatgaacaggaaatgacttactgcaacaaagatttagttcacatcgtctacagt base pairs cagtcatatcgactttaggagctacttgtcctttactgaatgacgttgtttctaaatcaagtgtagcagatgtca 751 to 825 ctttcaaaagacatggggttaccaggatttagagtcggaatcatatattcttttaacgacgatgtcgttaattgt base pairs gaaagttttctgtaccccaatggtcctaaatctcagccttagtatataagaaaattgctgctacagcaattaaca 826 to 900 gctagaaaaatgtcgagtttcggtttagtatctacacaaacgcaatattttttagcggcaatgctatcggacgaa base pairs cgatctttttacagctcaaagccaaatcatagatgtgtttgcgttataaaaaatcgccgttacgatagcctgctt 901 to 975 aaattcgtcgataattttctaagagaaagcgcgatgaggttaggtaaaaggcacaaacattttactaatggactt base pairs tttaagcagctattaaaagattctctttcgcgctactccaatccattttccgtgtttgtaaaatgattacctgaa 976 to 1050 gaagtagtgggaattaaatgcttgaaaaataatgcggggcttttttgttggatggatttgcgtccacttttaagg base pairs cttcatcacccttaatttacgaactttttattacgccccgaaaaaacaacctacctaaacgcaggtgaaaattcc 1051 to 1125 HindIII gaatcgactttcgatagcgaaatgtcgttatggagagttattataaacgatgttaagcttaacgtctcgcctgga base pairs cttagctgaaagctatcgctttacagcaatacctctcaataatatttgctacaattcgaattgcagagcggacct 1126 to 1200 tcttcgtttgaatgtcaagagccagggtggttccgagtttgttttgcaaatatggatgatggaacggttgatatt base pairs agaagcaaacttacagttctcggtcccaccaaggctcaaacaaaacgtttatacctactaccttgccaactataa 1201 to 1275 gcgctcgcgaggattcggaggttcgtaggtgttgagaaaagtggagataaatcgagttcgatggaaaagaagcaa base pairs cgcgagcgctcctaagcctccaagcatccacaactcttttcacctctatttagctcaagctaccttttcttcgtt 1276 to 1350 caatggaagaagaataatttgagacttagtttttcgaaaagaatgtatgatgaaagtgttttgtcaccactttcg base pairs gttaccttcttcttattaaactctgaatcaaaaagcttttcttacatactactttcacaaaacagtggtgaaagc 1351 to 1425 tcacctattcctccctcaccattagttcgttaa base pairs agtggataaggagggagtggtaatcaagcaatt 1426 to 1458 Table by Enzyme Name Enzyme No. Positions Recognition name cuts of sites sequence BamHI 1 108 g/gatcc EcoRI 1 289 g/aattc HindIII 1 1180 a/agctt Every enzyme analyzed cuts this sequence. Did your creating of a new BamH1 restriction site in your gene change the primary Amino acid sequence of your protein? Yes AMINO ACID SEQUENCE OF WILD TYPE/ORIGINAL GENE: Open reading frame 1 >EMBOSS_001_1 MGFEIAKTNSILSKLATNEEHGENSPYFDGWKAYDSDPFHPLKNPNGVIQMGLAENQLCL DLIEDWIKRNPKGSICSEGIKSFKAIANFQDYHGLPEFRKAIAKFMEKTRGGRVRFDPER VVMVGGATGANETIIFCLADPGDAFLVPSPYYPAFNRDLRWRTGVQLIPIHCESSNNFKI TSKAVKEAYENAQKSNIKVKGLILTNPSNPLGTTLDKDTLKSVLSFTNQHNIHLVCDEIY AATVFDTPQFVSIAEILDEQEMTYCNKDLVHIVYSLSKDMGLPGFRVGIIYSFNDDVVNC ARKMSSFGLVSTQTQYFLAAMLSDEKFVDNFLRESAMRLGKRHKHFTNGLEVVGIKCLKN NAGLFCWMDLRPLLRESTFDSEMSLWRVIINDVKLNVSPGSSFECQEPGWFRVCFANMDD GTVDIALARIRRFVGVEKSGDKSSSMEKKQQWKKNNLRLSFSKRMYDESVLSPLSSPIPP SPLVR AMINO ACID SEQUENCE OF THE MUTATED GENE: Open reading frame 1 >EMBOSS_001_1 MGFEIAKTNSILSKLATNEEHGENSPYFDGWKAYDRDPFHPLKNPNGVIQMGLAENQLCL DLIEDWIKRNPKGSICSEGIKSFKAIANFQDYHGLPEFRKAIAKFMEKTRGGRVRFDPER VVMVGGATGANETIIFCLADPGDAFLVPSPYYPAFNRDLRWRTGVQLIPIHCESSNNFKI TSKAVKEAYENAQKSNIKVKGLILTNPSNPLGTTLDKDTLKSVLSFTNQHNIHLVCDEIY AATVFDTPQFVSIAEILDEQEMTYCNKDLVHIVYSLSKDMGLPGFRVGIIYSFNDDVVNC ARKMSSFGLVSTQTQYFLAAMLSDEKFVDNFLRESAMRLGKRHKHFTNGLEVVGIKCLKN NAGLFCWMDLRPLLRESTFDSEMSLWRVIINDVKLNVSPGSSFECQEPGWFRVCFANMDD GTVDIALARIRRFVGVEKSGDKSSSMEKKQQWKKNNLRLSFSKRMYDESVLSPLSSPIPP SPLVR In creating a new restriction site for BamHI, the primary amino acid sequence of the protein changes. The original amino acid was Serine at position 36 which was substituted by Arginine after the mutation took place, (Ser36Arg). How many amino acid residues are in the protein? The number of amino acid residues in the protein is 485. What is the molecular weight of the protein? Its molecular weight is 54690.6 ProtParam 10 20 30 40 50 60 MGFEIAKTNS ILSKLATNEE HGENSPYFDG WKAYDSDPFH PLKNPNGVIQ MGLAENQLCL 70 80 90 100 110 120 DLIEDWIKRN PKGSICSEGI KSFKAIANFQ DYHGLPEFRK AIAKFMEKTR GGRVRFDPER 130 140 150 160 170 180 VVMVGGATGA NETIIFCLAD PGDAFLVPSP YYPAFNRDLR WRTGVQLIPI HCESSNNFKI 190 200 210 220 230 240 TSKAVKEAYE NAQKSNIKVK GLILTNPSNP LGTTLDKDTL KSVLSFTNQH NIHLVCDEIY 250 260 270 280 290 300 AATVFDTPQF VSIAEILDEQ EMTYCNKDLV HIVYSLSKDM GLPGFRVGII YSFNDDVVNC 310 320 330 340 350 360 ARKMSSFGLV STQTQYFLAA MLSDEKFVDN FLRESAMRLG KRHKHFTNGL EVVGIKCLKN 370 380 390 400 410 420 NAGLFCWMDL RPLLRESTFD SEMSLWRVII NDVKLNVSPG SSFECQEPGW FRVCFANMDD 430 440 450 460 470 480 GTVDIALARI RRFVGVEKSG DKSSSMEKKQ QWKKNNLRLS FSKRMYDESV LSPLSSPIPP SPLVR Number of amino acids: 485 Molecular weight: 54690.6 Theoretical pI: 7.92 Top of Form Amino acid composition:  Ala (A) 27 5.6% Arg (R) 24 4.9% Asn (N) 30 6.2% Asp (D) 29 6.0% Cys (C) 11 2.3% Gln (Q) 13 2.7% Glu (E) 28 5.8% Gly (G) 30 6.2% His (H) 9 1.9% Ile (I) 29 6.0% Leu (L) 42 8.7% Lys (K) 35 7.2% Met (M) 14 2.9% Phe (F) 29 6.0% Pro (P) 24 4.9% Ser (S) 41 8.5% Thr (T) 20 4.1% Trp (W) 7 1.4% Tyr (Y) 12 2.5% Val (V) 31 6.4% Pyl (O) 0 0.0% Sec (U) 0 0.0% (B) 0 0.0% (Z) 0 0.0% (X) 0 0.0% Bottom of Form Total number of negatively charged residues (Asp + Glu): 57 Total number of positively charged residues (Arg + Lys): 59 Atomic composition: Carbon C 2443 Hydrogen H 3816 Nitrogen N 660 Oxygen O 716 Sulfur S 25 Formula: C2443H3816N660O716S25 Total number of atoms: 7660 Extinction coefficients: Extinction coefficients are in units of M-1 cm-1, at 280 nm measured in water. Ext. coefficient 57005 Abs 0.1% (=1 g/l) 1.042, assuming all pairs of Cys residues form cystines Ext. coefficient 56380 Abs 0.1% (=1 g/l) 1.031, assuming all Cys residues are reduced Estimated half-life: The N-terminal of the sequence considered is M (Met). The estimated half-life is: 30 hours (mammalian reticulocytes, in vitro). >20 hours (yeast, in vivo). >10 hours (Escherichia coli, in vivo). Instability index: The instability index (II) is computed to be 42.67 This classifies the protein as unstable. Aliphatic index: 81.20 Grand average of hydropathicity (GRAVY): -0.285 What is the most abundant amino acid found in the protein? What is the least abundant amino acid found in the protein? The most abundant amino acid found in the protein is Leucine while the least abundant is Tryptophan. 1-Aminocyclopropane-1-Carboxylate Synthase belongs to class 1 aminotransferases and plays a critical role in the ethylene biosynthetic pathway in fruit development and ripening. This protein catalyzes the conversion of S-adenosylmethionine to ACC. This enzyme acts as a homodimer and one of its major co-factors is pridoxal-5-phosphate.In this pathway, regulation is majorly dependent on both positive and negative feedback mechanisms. Specifically, the positive feedback control mechanism is essential in the ripening of fruits and senescence of flowers. In tomato (Lycopersicon esculentum), there is an upsurge in ethylene production after exposure to exogenous ethylene. This in turn activates the activity of 1-Aminocyclopropane -1- Carboxylate Synthase. The role of 1-Aminocyclopropane-1-Carboxylate Synthase with regard to ripening of fruits was first demonstrated in the year 1991. In this experiment, its role was fully understood in transgenic tomato plants that expressed the protein antisense Ribonucleic acid (RNA). Further experimental studies involving mutagenesis and crystallization of the enzyme, three dimensional structures as well as amino acid residues have been of core importance in understanding its role. ACC synthase is beneficial in a number of ways including induction of apoptosis by oxidative stress, response to both external stimulus and ethylene. ACC synthase is important in cellular response to iron ions. Additionally, the protein plays a vital role in response to wounding, mechanical stimulus as well as jasmonic acid. This protein is expressed during various developmental and growth stages such as anthesis, leaf senescence, bilateral and globular stage. Within the plant cell, at molecular level, this enzyme is important in protein binding and is expressed in the flowers, leaf apex, embryo, stem and the leaves (Alexander and Don 2032). Control of ethylene production is fundamental aspect in plant biotechnology. This is because overproduction of ethylene is capable of delaying senescence and over maturation. As a result of these activities due to chemical changes, there has been a gradual occurrence of loss in storage of vegetables and fruits. Thus, there is the need for the design of detailed structural and biochemical data in combating the activity of this enzyme in terms of its inhibition. Such a procedure can be of great agricultural benefits to farmers which ultimately translate to food security in many nations that depend on agriculture. This protein is a cytosolic enzyme and catalyzes the rate limiting reaction in ethylene biosynthesis. It is difficult to purify this enzyme due to the fact that it is present in low quantities and also because of its labile nature. In the recent past, molecular cloning or transformation and functional gene expression of the protein in various cells have enabled the structural and biochemical studies. Molecular cloning of the complementary DNA and genomic sequences of this gene has been done from various plants. The genes emanating from these sequences show that ACC synthase is encoded by members of a divergent multi locus gene group. Each gene in the sequence of this protein is regulated in differential arrays with factors such as environmental and developmental playing a crucial role. There seems to be an emerging shift in studies targeting the regulation of ACC synthase isoforms. Such relevant and eye opening experiments have been undertaken on Arabidopsis. In this organism, Arabidopsis, characterization of this gene has led to the identification of seven sub genes. One specific gene is the acs2 which is induced by cycloheximide wounding or injury. This particular gene in the protein is shown to have a negative feedback inhibition mechanism on ethylene biosynthesis. Another isoform of the gene is ACS4 which is induced in seedlings by compounds such as indoleacetic acid, cycloheximide and injury. ACS5 is triggered by lithium chloride and lower concentration of shoot elongation factors called cytokinnins in seedlings. ACS6 is specifically induced by treatment with compounds that contain cyanide or exposure to ozone. However, this isoform can also be triggered by cycloheximide and indole acetic acid. ACS10 has been identified in the early developmental stages in Arabidopsis and is postulated to promote flowering. From the studies, cycloheximide has been shown to induce ACS isoforms and thus the implication is that short-lived and negatively regulated ACS transcripts are formed. In a particular study, potent inhibitors of the protein have been identified and one such inhibitor is cycloheximide. The use of biotechnological applications in reducing ethylene production has continued to reap enormous benefits hence enhancing fruit ripening and prolonging the shelf lives of the plants. Works Cited Alexander, Lucille and Don Grierson. "Ethylene biosynthesis and action in tomato: a model for climacteric fruit ripening." Journal of Experimental Botany (2002): 2039-2055. Arshad, Muhammad and William Frankenberger. "Ethylene in Plant Physiology." Journal of Biomedical and Life Sciences (2002): 11-50. Jakubowicz, Malgorzata and Jan Sadowski. "1-Aminocyclopropane-1-carboxylate synthase — genes and expression." Acta Physiologiae Plantarum (2002): 459-478. Ski, Jan and Ewa Ska. "Manipulation of ethylene biosynthesis ." Acta Physiologiae Plantarum (2001): 213-220. Read More