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Expression Mapping of Recombinant ISA1 Genes in Arabidopsis thaliana and Maize

This study focuses on the expression mapping of the recombinant ISA1 genes, AtISA1-FL and ZmISA1-FL, in Arabidopsis thaliana and maize. Full-length cDNA sequences of the ISA1 gene from both species are introduced using the shuttle vector pMDC32. This approach allows for the investigation of gene function and expression patterns in different plant species, aiming to enhance our understanding of carbohydrate metabolism. This research contributes valuable data for future plant biotechnology applications.

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Expression Mapping of Recombinant ISA1 Genes in Arabidopsis thaliana and Maize

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AtISA1-FL ATGGATGCAATCAAATGCAGTTCCAGTTTCCTCCATCACACCAAGCTCAACACTCTCTTCTCTAATCACACCTTTCCTAAAATCTCCGCTCCCAATTTCAAGCCCCTCTTCCGTCCGATTTCGATTTCCGCAAAGGACAGAAGAAGCAACGAAGCTGAAAATATCGCCGTAGTAGAGAAACCTCTTAAATCAGATAGATTTTTTATCTCCGATGGACTTCCTTCTCCATTCGGACCCACCGTCAGAGACGACGGCGTCAATTTCTCTGTTTACTCTACAAACTCCGTTTCCGCTACCATCTGCTTGATTTCTCTCTCCGATCTCCGTCAGAACAAAGTGACGGAGGAGATTCAGCTTGATCCATCAAGGAATAGAACTGGCCATGTTTGGCATGTGTTCTTGAGAGGAGATTTCAAAGATATGTTGTATGGTTATAGATTTGATGGCAAGTTTTCTCCTGAAGAAGGTCATTATTATGATTCCTCCAACATTTTATTGGATCCTTACGCAAAGGCAATTATAAGCAGAGATGAGTTTGGAGTTTTGGGACCTGATGATAATTGTTGGCCTCAAATGGCGTGTATGGTACCCACTCGTGAGGAAGAGTTTGATTGGGAAGGGGATATGCATCTGAAGCTTCCACAGAAAGATCTTGTTATATATGAAATGCATGTGCGAGGTTTTACAAGGCATGAGTCTAGTAAAATTGAATTCCCTGGCACATACCAGGGTGTTGCAGAGAAGCTTGACCATTTGAAGGAGCTTGGGATAAATTGTATAGAATTAATGCCATGTCACGAGTTTAATGAGCTGGAGTATTACAGCTACAATACGATTTTGGGAGACCACAGGGTAAATTTTTGGGGTTACTCTACCATTGGGTTCTTCTCGCCCATGATCAGATACGCATCAGCAAGCTCTAACAATTTTGCTGGACGAGCCATAAATGAATTCAAAATTCTTGTTAAGGAGGCACATAAACGAGGAATTGAGGTAATCATGGATGTCGTCTTGAACCACACAGCCGAAGGGAATGAAAAAGGGCCCATTTTCTCATTTAGAGGAGTTGATAACAGTGTCTATTACATGCTTGCTCCAAAGGGCGAGTTCTATAATTATTCAGGCTGTGGTAATACATTCAACTGCAATCATCCTGTGGTGCGTCAATTCATATTGGATTGCCTGAGATATTGGGTTACAGAAATGCATGTTGACGGCTTCCGCTTTGATCTTGGTTCAATCATGTCAAGGAGCAGCAGCCTTTGGGATGCAGCCAATGTTTACGGGGCTGATGTAGAAGGTGACTTGCTCACAACTGGTACTCCTATTAGCTGCCCTCCAGTAATTGACATGATAAGTAATGATCCAATACTCCGCGGTGTTAAGCTAATAGCTGAAGCATGGGATGCGGGTGGGCTGTACCAAGTTGGCATGTTTCCACACTGGGGTATTTGGTCTGAGTGGAATGGAAAGTTTCGGGATGTTGTGAGACAGTTCATAAAAGGCACCGATGGCTTTTCTGGTGCTTTTGCTGAATGTCTCTGTGGAAGCCCAAATCTGTACCAGGGAGGTAGGAAACCTTGGCACAGCATCAATTTTATATGTGCGCATGATGGTTTTACGTTGGCAGATTTAGTAACTTACAACAATAAGAATAACTTGGCAAATGGAGAAGAGAATAATGATGGAGAGAATCACAATTACAGCTGGAACTGTGGAGAGGAGGGAGACTTTGCGAGTATCTCGGTGAAGAGACTAAGGAAACGACAGATGCGGAATTTCTTTGTTTCCCTCATGGTTTCCCAAGGTGTCCCAATGATTTACATGGGAGATGAATATGGCCATACTAAAGGGGGAAACAACAACACGTATTGCCATGACAACTATATGAACTATTTTCGGTGGGATAAAAAGGAAGAAGCACATTCTGACTTCTTCAGATTCTGCCGTATTCTTATCAAGTTTCGTGATGAATGTGAATCACTTGGCTTGAATGATTTCCCAACAGCAAAGCGTCTGCAGTGGCATGGTCTTGCTCCTGAGATCCCAAATTGGTCTGAAACAAGTCGATTTGTTGCATTTTCACTGGTCGATTCTGTGAAGAAAGAAATCTATGTGGCCTTCAACACCAGTCATTTAGCCACACTTGTTTCGCTACCGAATAGGCCAGGATACCGATGGGAGCCATTTGTAGACACGAGCAAACCGAGCCCTTACGACTGCATAACACCGGATCTCCCAGAGAGAGAAACGGCGATGAAGCAGTATAGGCACTTCTTAGATGCAAATGTGTACCCAATGCTCAGTTACTCATCCATCATTCTCCTTCTTTCACCAATTAAAGACCCCTGA ZmISA1-FL ATGGATGCAATCAAATGCAGTTCCAGTTTCCTCCATCACACCAAGCTCAACACTCTCTTCTCTAATCACACCTTTCCTAAAATCTCCGCTCCCAATTTCAAGCCCCTCTTCCGTCCGATTTCGATTTCCGTGGCCGAGGCGGTGCAGGCGGAGGAGGACGACGACGACGACGACGAGGAGGTGGCCGAGGAGAGGTTCGCGCTGGGCGGCGCGTGCCGGGTGCTCGCGGGAATGCCCGCGCCGCTCGGCGCCACCGCGCTCCGCGGCGGTGTCAACTTCGCCGTCTACTCCAGCGGTGCCTCCGCCGCGTCGCTGTGCCTCTTCGCTCCCGGCGACCTCAAGGCGGATAGGGTGACCGAGGAGGTGCCCCTCGATCCCCTGCTCAACCGAACGGGAAACGTGTGGCACGTGTTCATCCACGGGGACCAGCTGCACGGCATGCTCTACGGATACAGGTTCGATGGCGTGTTCGCCCCTGAGCGCGGACAGTACTACGATGTGTCCAACGTTGTGGTGGATCCATACGCTAAGGCAGTGGTAAGCCGAGGTGAATATGGTGTGCCTGCGCCTGGTGGTAGTTGTTGGCCTCAAATGGCTGGTATGATCCCTCTTCCCTATAATAAGTTTGATTGGCAAGGTGACCTACCCCTTGGGTACCATCAGAAGGACCTTGTCATATATGAAATGCATTTGCGTGGATTCACAAAGCACAACTCAAGCAAGACAAAACACCCAGGAACTTACATTGGTGCTGTGTCAAAGCTTGACCATCTAAAGGAACTTGGAGTGAACTGTATAGAGCTAATGCCCTGCCATGAGTTCAATGAGCTAGAGTACTTCAGCTCCTCTTCGAAGATGAACTTCTGGGGATATTCCACAATAAATTTTTTCTCACCAATGGCAAGATATTCTTCAAGTGGCATAAGAGACTCTGGATGTGGTGCCATAAATGAATTTAAAGCTTTTGTAAGGGAGGCCCACAAACGGGGAATTGAGGTGATCATGGATGTTGTCTTCAATCATACAGCTGAAGGTAATGAGAAAGGCCCAATATTATCCTTTAGGGGGATAGATAATAGTACATACTACATGCTTGCACCTAAGGGAGAGTTTTATAATTATTCTGGTTGTGGAAATACCTTCAATTGTAATCATCCTGTAGTCCGTGAATTTATAGTGGATTGCTTGAGATACTGGGTAACAGAAATGCATGTTGATGGTTTTCGTTTTGACCTTGCATCTATACTGACCAGAGGATGCAGTCTATGGGATCCAGTTAATGTGTATGGAAGTCCAATGGAAGGTGACATGATTACGACAGGGACACCTCTTGTTGCCCCACCACTTATTGACATGATTAGCAATGACCCAATTCTTGGAAATGTCAAGCTCATTGCTGAAGCATGGGATGCAGGAGGTCTCTATCAAGTTGGTCAGTTTCCTCACTGGAACGTTTGGTCAGAGTGGAATGGAAAGTATCGCGATACCGTGCGTCAGTTCATCAAAGGCACAGATGGATTTGCTGGTGCTTTTGCTGAATGCCTATGTGGAAGTCCACAGTTATACCAGGCAGGGGGGAGGAAGCCTTGGCACAGTATCAACTTTGTATGTGCACACGATGGATTTACACTGGCTGATTTGGTCACATACAATAGCAAGTACAACTTGTCAAATGGTGAGGACAACAGAGATGGGGAAAATCATAATCTTAGCTGGAATTGTGGGGAGGAAGGAGAATTTGCAAGTCTGTCAGTCCGAAGATTAAGGAAGAGGCAAATGCGCAATTTCTTTGTTTGTCTTATGGTTTCTCAGGGAGTTCCAATGTTCTACATGGGCGATGAATATGGTCACACAAAGGGAGGGAACAACAATACGTACTGCCATGACCATTATGTCAATTATTTCCGTTGGGATAAGAAGGAAGAACAATCCTCTGATTTGTACAGATTCTGCCGTCTCATGACCAAATTCCGCAAAGAATGTGAATCTCTTGGCCTTGAGGACTTCCCGACTTCAGAACGGTTGAAATGGCACGGTCATCAGCCCGAGAAGCCTGACTGGTCAGAGGCAAGCCGATTCGTTGCCTTCACCATGAAGGACGAAACCAAAGGCGAGATCTACGTGGCCTTCAACACCAGTCACCTTCCGGTGGTTGTTGGGCTTCCAGAGCGCTCTGGGTTCCGATGGGAGCCGGTGGTGGACACCGGCAAGGAGGCACCATATGACTTCCTCACCGATGGCCTGCCAGATCGTGCTGTCACCGTCTACCAGTTCTCTCATTTCCTCAACTCCAATCTCTATCCTATGCTCAGCTACTCCTCCATCATCCTTGTATTGCGCCCTGATGTCTGA Supplemental Figure S3: Map of the shuttle vector pMDC32 used for expression of the recombinant genes AtISA1-FL and and ZmISA1-FL into Arabidopsis thaliana. Arabidopsis (AtISA1-FL) and maize (ZmISA1-FL) full length cDNA sequences of ISA1 gene (depicted above) were introduced between the right (RB) and left (LB) borders of the modified T-DNA and placed under the control of the strong and constitutive 2x35S promoter. The NosT terminator sequence was placed downstream of ISA1 cDNA. KanR is the selection marker for E. coli and Agrobacterium tumefaciens conferring resistance to kanamycin. HygR is the selection gene that confers resistance to hygromycin B to the transformed plants.

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