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Cloning and Functional Analysis of APETALA1 Promoter

  • Received Date: 2012-08-24
  • APETALA1 is an important MADS box gene involved in the regulatory pathway of flowering. The expression of the AP1 decreases dramatically in the Abnormal Flower Development Line (AFDL) of Arabidopsis. AFDL is absent of petals and with a stacked inflorescence meristems. In order to analysis the matically in the Abnormal Flower Development Line (AFDL) of Arabidopsis. AFDL is absent of petals and with a stacked inflorescence meristems. In order to analysis the regulation of the promoter of AP1, the -3 579 bp promoter region of AP1 was cloned from AFDL. The functional element predication indicated that three CArG boxes, the binding sites for the MADS regulator, were located in the promoter (the three boxes were identified by progressive number as CArG1, CArG2, CArG3 form). 5 vectors were constructed with the promoters of AP1 to drive GUS in wild-type, which with different length and number of the CArG boxes. The sequence of AP1 in AFDL was the same as that of wild-type, that indicated that the significant down-regulation of AP1 was not associated with the CArG boxes as well as its sequenced; And the GUS staining of the transformed plants showed that the CArG1 was the binding site for positively acting factor(s) during the early stage and the later stage of the flower development, and the deletion of the CArG2 and the mutations in CArG3 resulted in an increase in the level of reporter gene activity during the early or later floral stages, suggesting that CArG2 and CArG3 are the binding sites for negatively regulators. Therefore the three CArG boxes in the promoter of AP1 are functional on the regulation the AP1 expression rather than determinant expression of AP1. The ectopic GUS expression in the whorl 4 indicated that there were unidentified elements that played a role on the expression specificity outside the 0-3 579 bp upstream of the transcriptional start site. In addition, there may be some elements located in the region from -3 579 bp to -1 752 bp but the region from -1 752 bp to -1 359 bp was nonfunctional on the regulation of AP1 except the CArG2 box.
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Cloning and Functional Analysis of APETALA1 Promoter

  • 1. Research Institute of Forestry, State Key Laboratory of Tree Genetics and Breeding, Chinese Academy of Forestry, Beijing 100091, China

Abstract: APETALA1 is an important MADS box gene involved in the regulatory pathway of flowering. The expression of the AP1 decreases dramatically in the Abnormal Flower Development Line (AFDL) of Arabidopsis. AFDL is absent of petals and with a stacked inflorescence meristems. In order to analysis the matically in the Abnormal Flower Development Line (AFDL) of Arabidopsis. AFDL is absent of petals and with a stacked inflorescence meristems. In order to analysis the regulation of the promoter of AP1, the -3 579 bp promoter region of AP1 was cloned from AFDL. The functional element predication indicated that three CArG boxes, the binding sites for the MADS regulator, were located in the promoter (the three boxes were identified by progressive number as CArG1, CArG2, CArG3 form). 5 vectors were constructed with the promoters of AP1 to drive GUS in wild-type, which with different length and number of the CArG boxes. The sequence of AP1 in AFDL was the same as that of wild-type, that indicated that the significant down-regulation of AP1 was not associated with the CArG boxes as well as its sequenced; And the GUS staining of the transformed plants showed that the CArG1 was the binding site for positively acting factor(s) during the early stage and the later stage of the flower development, and the deletion of the CArG2 and the mutations in CArG3 resulted in an increase in the level of reporter gene activity during the early or later floral stages, suggesting that CArG2 and CArG3 are the binding sites for negatively regulators. Therefore the three CArG boxes in the promoter of AP1 are functional on the regulation the AP1 expression rather than determinant expression of AP1. The ectopic GUS expression in the whorl 4 indicated that there were unidentified elements that played a role on the expression specificity outside the 0-3 579 bp upstream of the transcriptional start site. In addition, there may be some elements located in the region from -3 579 bp to -1 752 bp but the region from -1 752 bp to -1 359 bp was nonfunctional on the regulation of AP1 except the CArG2 box.

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