Диссертация (1145914), страница 21
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Приложения1.Нуклеотидная последовательность, составленная по результатам секвенирования плазмиды pPIC9- АОХ1-PHO5 с использованием праймеров: PHO5F (CGGGATCCCGAGATTACCAA), PHO5R (CGGAATTCCAAAACTATTGT),5'AOX1 (GACTGGTTCCAATTGACAAGC) и 3'AOX1 (GCAAATGGCATTCTGACATCC):AACTAATTATTCGAAGGATCCCGAGATTACCAATGTTTAAATCTGTTGTTTATTCAATTTTAGCCGCTTCTTTGGCCAATGCAGGTACCATTCCCTTAGGCAAACTAGCCGATGTCGACAAGATTGGTACCCAAAAAGATATCTTCCCATTTTTGGGTGGTGCCGGACCATACTACTCTTTCCCTGGCGACTATGGTATTTCTCGTGATTTGCCTGAAGGTTGTGAAATGAAGCAACTGCAAATGGTTGGTAGACATGGTGAAAGATACCCTACTGTCAGTCTGGCTAAGACTATCAAGAGTACATGGTATAAGTTGAGCAATTACACTCGTCAATTCAACGGCTCATTGTCATTCTTGAACGATGATTACGAGTTTTTCATCCGTGATGACGATGATTTGGAAATGGAAACCACTTTTGCCAACTCGGACGATGTTTTGAACCCATACACTGGTGAAATGAACGCCAAGAGACATGCTCGTGACTTCTTGGCTCAATACGGTTACATGGTCGAAAACCAAACCAGTTTCGCCGTTTTTACCTCTAATTCTAAGAGATGTCATGACACTGCTCAATATTTCATTGATGGTTTAGGTGACCAATTCAACATCACCTTGCAGACTGTCAGTGAAGCTGAATCCGCTGGTGCCAACACTTTGAGTGCTTGTAACTCATGTCCTGCTTGGGACTACGATGCCAATGATGACATTGTAAATGAATACGACACAACCTACTTGGATGACATTGCCAAGAGATTGAACAAGGAAAACAAGGGTTTGAACTTGACCTCAACTGACGCTAGTACTTTATTCTCGTGGTGTGCATTTGAAGTGAACGCTAAAGGTTACAGTGATGTCTGTGATATTTTCACCAAGGATGAATTAGTCCATTACTCCTACTACCAAGACTTGCACACTTATTACCATGAGGGTCCAGGTTACGACATTATCAAGTCTGTCGGTTCCAACTTGTTCAATGCCTCAGTCAAATTATTAAAGCAAAGTGAGATTCAAGACCAAAAGGTTTGGTTGAGTTTTACCCACGATACCGATATCCTAAACTTTTTG143ACCACCGCTGGTATAATTGACGACAAAAACAACTTAACTGCCGAATACGTTCCATTCATGGGCAACACTTTCCACAGATCCTGGTACGTTCCTCAAGGTGCTCGTGTCTACACCGAAAAATTCCAATGTTCTAACGACACCTACGTCAGATACGTCATTAACGATGCTGTTGTTCCAATTGAAACCTGTTCCACTGGTCCAGGGTTCTCTTGTGAAATCAATGACTTCTACGACTATGCTGAAAAGAGAGTAGCCGGTACTGACTTCCTAAAGGTCTGTAACGTCAGCAGCGTCAGTAACTCTACTGAATTGACCTTCTACTGGGACTGGAACACTACTCATTACAACGCCAGTCTATTGAGACAATAGTTTTGGAATTCCCTAGGGCGGCCGCGAATTAATTCGCCTTAGACATGACTGTTCCTCCGAA – часть последовательности промотора гена АОХ1,GGATCC – сайт рестрикции BamHI,GAATTC – сайт рестрикции EcoRI,CAGA – последовательность гена КФ PHO5,CTAG – часть последовательности 3' гена АОХ1.2.PHO5Нуклеотидная последовательность, составленная по результатам секвенирования плазмиды pPIC9-АОХ2сиспользованиемпраймеров:PAOX2F(TTCGACGTCCTGCCCTCTCC),PAOX2R(TTCGGATCCTTTTCTCAGTTGATTTG), PHO5Seq (GACATCGGCTAGTTTGC).TTGAAAAAGTTGACCTTCTAAGATCTCAAAAACCTAAGTACTTCATTTGAATATAACTCTGCACCTAAATTTACACCTAACTCTCTATCTAGGCTCTAGATTTGATAGATTCTATAGCCTTTGGTTTGTTATAGTGTTCACCAACTGGATGTCCTAACGAAATGGTTCTGTGGTCTAGTTGGTTATGGCATATGCTTAACACGCATAACGTCCCCAGTTCGATCCTGGGCA144GAATCATTATTTTTTGACCGAATTCTTTTTTTCAGACCATATGACCGGTCCATCTTCTACGGGGGGATTATCTATGCTTTGACCTCTATCTTGATTCTTTTATGATTCAAATCACTTTTACGTTATTTATTACTTACTGGTTATTTACTTAGCGCCTTTTCTGAAAAACATTTACTAAAAATCATACATCGGCACTCTCAAACACGACAGATTGTGATCAAGAAGCAGAGACAATCACCACTAAGGTTGCACATTTGAGCCAGTAGGCTCCTAATAGAGGTTCGATACTTATTTTGATAATACGACATATTGTCTTACCTCTGAATGTGTCAATACTCTCTCGTTCTTCGTCTCGTCAGCTAAAAATATAACACTTCGAGTAAGATACGCCCAATTGAAGGCTACGAGATACCAGACTATCACTAGTAGAACTTTGACATCTGCTAAAGCAGATCAAATATCCATTTATCCAGAATCAATTACCTTCCTTTAGCTTGTCGAAGGCATGAAAAAGCTA……TTAAGCATAGATTGATGGAGGGTGTATGGCACTTGGCGGCTGCATTAGAGTTTGAAACTATGGGGTAATACATCACATCCGGAACTGATCCGACTCCGAGATCATATGCAAAGCACGTGATGTACCCCGTAAACTGCTCGGATTATCGTTGCAATTCATCGTCTTAAACAGTACAAGAAACTTTATTCATGGGTCATTGGACTCTGATGAGGGGCACATTTCCCCAATGATTTTTTGGGAAAGAAAGCCGTAAGAGGACAGTTAAGCGAAAGAGACAAGACAACGAACAGCAAAAGTGACAGCTGTCAGCTACCTAGTGGACAGTTGGGAGTTTCCAATTGGTTGGTTTTGAATTTTTACCCATGTTGAGTTGTCCTTGCTTCTCCTTGCAAACAATGCAAGTTGATAAGACATCACCTTCCAAGATAGGCTATTTTTGTCGCATAAATTTTTGTCTCGGAGTGAAAACCCCTTTTATGTGAACAGATTACAGAAGCGTCCTACCCTTCACCGGTTGAGATGGGGAGAAAATTAAGCGATGAGGAGACGATTATTGGTATAAAAGAAGCAACCAAAATCCCTTATTGTCCTTTTCTGATCAGCATCAAAGAATATTGTCTTAAAACGGGCTTTTAACTACATTGTTCTTACACATTGCAAACCTCTTCCTTCTATTTCGGATCAACTGTATTGACTACATTGATCTTTTTTAACGAAGTTTACGACTTACTAAATCCCCACAAACAAATCAACTGAGAAAAGGATCCCGAGATTACCAATGTTTAA145CGAA – последовательность промотора гена АОХ2,GGATCC – сайт рестрикции BamHI,CAGA – часть последовательности гена КФ PHO5.3.Нуклеотидная последовательность, полученная при секвенировании плазмиды pPIC9-ΔSacI-PHO5 сиспользованием праймера: PIC9Seq (GGTTATTGTCTCATGAGC):AAAAGTGCCACCTGACGTAGCTCGCTCATTCCAATTCCTTCTATTAGGCTACTAACACCATGACTTTATTAGCCTGTCTATCCTGGCCCCCCTGGCGAGGTTCATGTTTGTTTATTTCCGAATGCAACAAGCTCCGCATTACACCCGAACATCACTCCAGATGAGGGCTTTCTGAGTGTGGGGTCAAATAGTTTCATGTTCCCCAAATGGCCCAAAACTGACAGTTTAAACGCTGTCTTGGAACCTAATATGACAAAAGCGTGATCTCATCCAAGATGAACTAAGTTTGGTTCGTTGAAATGCTAACGGCCAGTTGGTCAAAAAGAAACTTCCAAAAGTCGCCATACCGTTTGTCTTGTTTGGTATTGATTGACGAATGCTCAAAAATAATCTCATTAATGCTTAGCGCAGTCTCGAA – часть последовательности промотора гена АОХ1,GACGT – часть сайта рестрикции AatII,AGCTC – часть сайта рестрикции SacI,4.Нуклеотидная последовательность полученная при секвенировании плазмиды pPIC9-ΔPmeI-PHO5 сиспользованием праймера: PIC9Seq (GGTTATTGTCTCATGAGC):146CACCTGACGTAAACGCTGTCTTGGAACCTAATATGACAAAAGCGTGATCTCATCCAAGATGAACTAAGTTTGGTTCGTTGAAATGCTAACGGCCAGTTGGTCAAAAAGAAACTTCCAAAAGTCGCCATACCGTTTGTCTTGTTTGGTATTGATTGACGAATGCTCAAAAATAATCTCATTAATGCTTAGCGCAGTCTCTCTATCGCTTCTGAACCCCGGTGCACCTGTGCCGAAACGCAAATGGGGAAACACCCGCTTTTTGGATGATTATGCATTGTCTCCACATTGTATGCTTCCAAGATTCTGGTGGGAATACTGCTGATAGCCTAACGTTCATGATCAAAATTTAACTGTTCTAACCCCTACTTGACAGCAATATATAAACAGAAGGAAGCTGCCCTGTCTTAAACCTTTTTTTTTATCATCATTATTAGCTTACTTTCATAATTGCGACTGGTTCCAACGAA – часть последовательности промотора гена АОХ1,GACGT – часть сайта рестрикции AatII,AAAC – часть сайта рестрикции PmeI,5.Нуклеотидная последовательность полученная при секвенировании плазмиды pPIC9-ΔNsiI-PHO5 сиспользованием праймера: PIC9Seq (GGTTATTGTCTCATGAGC):CCTGACGTTGCATTGTCTCCACATTGTATGCTTCCAAGATTCTGGTGGGAATACTGCTGATAGCCTAACGTTCATGATCAAAATTTAACTGTTCTAACCCCTACTTGACAGCAATATATAAACAGAAGGAAGCTGCCCTGTCTTAAACCTTTTTTTTTATCATCATTATTAGCTTACTTTCATAATTGCGACTGGTTCCAATTGACAAGCTTTTGATTTTAACGACTTTTAACGACAACTTGAGAAGATCAAAAAACAACTAATTATTCGAAGGATCCCGAGATTACCAATGTTTAAATCTGTTGTTT147ATTCAATTTTAGCCGCTTCTTTGGCCAATGCAGGTACCATTCCCTTAGGCAAACTAGCCGATGTCGACAAGATTGGTACCCAAAAAGATATCTTCCGAA – часть последовательности промотора гена АОХ1,GACGT – часть сайта рестрикции AatII,TGCATT – часть сайта рестрикции NsiI,GGATCC – сайт рестрикции BamHI,CAGA – последовательность гена КФ PHO5,6.Нуклеотидная последовательность полученная при секвенировании плазмиды pPIC9-ΔEspI-PHO5 сиспользованием праймера: PIC9Seq (GGTTATTGTCTCATGAGC):AGTGCCACCTGACGTGCGCAGTCTCTCTATCGCTTCTGAACCCCGGTGCACCTGTGCCGAAACGCAAATGGGGAAACACCCGCTTTTTGGATGATTATGCATTGTCTCCACATTGTATGCTTCCAAGATTCTGGTGGGAATACTGCTGATAGCCTAACGTTCATGATCAAAATTTAACTGTTCTAACCCCTACTTGACAGCAATATATAAACAGAAGGAAGCTGCCCTGTCTTAAACCTTTTTTTTTATCATCATTATTAGCTTACTTTCATAATTGCGACTGGTTCCAATTGACAAGCTTTTGATTTTAACGACTTTTAACGACAACTTGAGAAGATCAAAAAACAACTAATTATTCGAAGGATCGAA – часть последовательности промотора гена АОХ1,GACGT – часть сайта рестрикции AatII,148TGCATT – часть сайта рестрикции EspI,GGATCC – часть сайта рестрикции BamHI,7.Нуклеотидная последовательность полученная при секвенировании плазмиды pPIC9-ΔA-PHO5 сиспользованием праймерa PHO5Seq (GACATCGGCTAGTTTGC):GTTTGGTTCGTTGAAATGCTAACGGCCAGTTGGTCAAAAAGAAACTTCCAAAAGTCGCCATACCGTTTGTCTTGTTTGGTATTGATTGACGAATGCTCAAAAATAATCTCATTAATGCTTAGCGCAGTCTCTCTATCGCTTCTGAACCCCGGTGCACCTGTGCCGAAACGCAAATTAGCCTAACGTTCATGATCAAAATTTAACTGTTCTAACCCCTACTTGACAGCAATATATAAACAGAAGGAAGCTGCCCTGTCTTAAACCTTTTTTTTTATCATCATTATTAGCTTACTTTCATAATTGCGACTGGTTCCAATTGACAAGCTTTTGATTTTAACGACTTTTAACGACAACTTGAGAAGATCAAAAAACAACTAATTATTCGAAGGATCCCGAGATTACCAATGTTTAAATCTGTTЗдесь и далее:CGAA – часть последовательности промотора гена АОХ1,TT – месторасположение делеции,GGATCC – сайт рестрикции BamHI,CAGA – часть последовательности гена КФ PHO5,1498.Нуклеотидная последовательность полученная при секвенировании плазмиды pPIC9-ΔВ-PHO5 сиспользованием праймерa PHO5Seq (GACATCGGCTAGTTTGC):GTTTGTTTATTTCCGAATGCAACAAGCTCCGCATTACACCCGAACATCACTCCAGATGAGGGCTTTCTGAGTGTGGGGTCAAATAGTTTCATGTTCCCCAAATGGCCCAAAACTGACAGTTTAAACGCTGTCTTGGAACCTAATATGACAAAAGCGTGATCTCATCCAAGATGAACTAAGTTTGGTTCGTTGAAATGCTAACGGCCAGTTGGTCAAAAAGAAACTTCCAAAAGTCGCCATACCGTTTGTCTTGTTTGGTATTGATTGACGAATGCTCAAAAATAATCTCATTAATGCTTAGCGCAGTCTCTCTATCGCTTCTGAACCCCGGTGCACCTGTGCCGAAACGCAAATGGGGAAACACCCGCTTTTTGGATGATTATGCATTGTCTCCACATTGTATGCTTCCAAGATTCTGGTGGGAATACTGTTCTAACCCCTACTTGACAGCAATATATAAACAGAAGGAAGCTGCCCTGTCTTAAACCTTTTTTTTTATCATCATTATTAGCTTACTTTCATAATTGCGACTGGTTCCAATTGACAAGCTTTTGATTTTAACGACTTTTAACGACAACTTGAGAAGATCAAAAAACAACTAATTATTCGAAGGATCCCGAGATTACCAATGTT9.Нуклеотидная последовательность полученная при секвенировании плазмиды pPIC9-ΔС-PHO5 сиспользованием праймерa PHO5Seq (GACATCGGCTAGTTTGC):CTCATTCCAATTCCTTCTATTAGGCTACTAACACCATGACTTTATTAGCCTGTCTATCCTGGCCCCCCTGGCGAGGTTCATGTTTGTTTATTTCCGAATGCAACAAGCTCCGCATTACACCCGAACATCACTCCAGATGAGGGCTTTCTGAGTGTGGGGTCAAATAGTTTCATGTTCCCCAAATGGCCCAAAACTGACAGTTTAAACGCTGTCTTGGAACCTAATATGACAAAAGCGTGATCTCATCCAAGATGAACTAAGTTTGGTTCGTTGAAATGCTAACGGCCAGTTGGTCAAAAAGAAACTTCCAAAAGTCGCCATACCGTTTGTCTTGTTTGGTATTGATTGACGAATGCTCAAAAATAATCTCATTAATGCTTAG150CGCAGTCTCTCTATCGCTTCTGAACCCCGGTGCACCTGTGCCGAAACGCAAATGGGGAAACACCCGCTTTTTGGATGATTATGCATTGTCTCCACATTGTATGCTTCCAAGATTCTGGTGGGAATACTGCTGATAGCCTAACGACAACTTGAGAAGATCAAAAAACAACTAATTATTCGAAGGATCCCGAGATTACCAATGTTTA10.Нуклеотидная последовательность полученная при секвенировании плазмиды pPIC9-ΔD-PHO5 сиспользованием праймерa PHO5Seq (GACATCGGCTAGTTTGC):CTAACACCATGACTTTATTAGCCTGTCTATCCTGGCCCCCCTGGCGAGGTTCATGTTTGTTTATTTCCGAATGCAACAAGCTCCGCATTACACCCGAACATCACTCCAGATGAGGGCTTTCTGAGTGTGGGGTCAAATAGTTTCATGTTCCCCAAATGGCCCAAAACTGACAGTTTAAACGCTGTCTTGGAACCTAATATGACAAAAGCGTGATCTCATCCAAGATGAACTAAGTTTGGTTCGTTGAAATGCTAACGGCCAGTTGGTCAAAAAGAAACTTCCAAAAGTCGCCATACCGTTTGTCTTGTTTGGTATTGATTGACGAATGCTCAAAAATAATCTCATTAATGCTTAGCGCAGTCTCTCTATCGCTTCTGAACCCCGGTGCACCTGTGCCGAAACGCAAATGGGGAAACACCCGCTTTTTGGATGATTATGCATTGTCTCCACATTGTATGCTTCCAAGATTCTGGTGGGAATACTGCTGATAGCCTAACGTTCATGATCAAAATTTAACTGTTCTAACCCCTACTTGACAGCAATATATAAACAGAAGGAAGCTGCCCTGTCTTAAACCTTTTTTTTTATCATCATTATTAGCTTACTTTCATAATTGCGACTGGATCCCGAGATTACCAATGTTTAAATCT11.
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