[ Objective ] This study aimed to investigate the expression of IPT gene directed by GmSARK promoter in Arabidopsis. ~ Method J IPT gene and C.m- SARK promoter were respectively cloned to construct plant expression ve...[ Objective ] This study aimed to investigate the expression of IPT gene directed by GmSARK promoter in Arabidopsis. ~ Method J IPT gene and C.m- SARK promoter were respectively cloned to construct plant expression vector and transform Arabidopsis. Tl transgenie plant lines were detected by PPT （ phosphino- thricin） herbicide selection and treated under drought and dark conditions for semi-quantitive RT-PCR analysis. E Result] GmSARK： ：IPT fusion gene was success- fully cloned and the plant expression vector p3301-GmSARK-IlYF was constructed. RT-PCR analysis showed that the target gene was expressed in T1 transgeuie plant lines at the mRNA level. [ Conclusion] This study laid the foundation for further investigating the roles of IPT gene directed by GmSARK promoter in plant stress resistance.展开更多
Fruit specific promoter (2A12) from Lycopersi-com esculentum and cDNA of isopentenyl-transferase (ipt) from Ti plasmid of Agrobacterium tumerfaciens C58 were cloned by PCR procedure respectively. Two plant expression ...Fruit specific promoter (2A12) from Lycopersi-com esculentum and cDNA of isopentenyl-transferase (ipt) from Ti plasmid of Agrobacterium tumerfaciens C58 were cloned by PCR procedure respectively. Two plant expression vectors with 2A12/gus or 2A12/ipt were respectively constructed. These two chimeric genes were transferred into tomato by Agrobacterium mediated procedure. The results of Southern hybridization showed that the fusion genes had been integrated into tomatoes. The result of gus histochemi-cal staining showed that 2A12 had high fruit specific expressive capability in transgenic tomato. The ipt expression resulted in accumulation of high level of cytokinins (CTKs) in fruit lead to developmental changes in fruits and seeds. The fruit of ipt transformed tomato had the hyperplastic placenta with very few seeds or even seedless. The shelf life of transgenic fruits elongated for 1-2 weeks. The ratio of fruit set, the dry weight of fruit and the crude protein content in fruit were increased, while展开更多
The isopenteryl transferase ( ipt ) gene from Agrobacterium tumefaciens (Smith et Townsend) Conn was driven under the tobacco TA29 promoter and introduced into tobacco (Nicotiana tabacum L.) plants by A. tumefaciens m...The isopenteryl transferase ( ipt ) gene from Agrobacterium tumefaciens (Smith et Townsend) Conn was driven under the tobacco TA29 promoter and introduced into tobacco (Nicotiana tabacum L.) plants by A. tumefaciens mediated transformation. PCR and Southern blot analysis confirmed that the ipt gene has integrated into the genomes of tobacco plants. The expression pattern of this chimeric TA29_ ipt gene in the transgenic plants was studied, and the endogenous cytokinin level in different organs was assayed by ELISA method. The results showed that the cytokinin content in the androecium of transgenic plants increased 3-4 times as compared with the control, and some changes of the fertility of the TA29_ ipt transgenic plants have been observed.展开更多
基金Supported by National Major Special Project of China on New Varieties Cultivation for Transgenic Organisms(NO.2010ZX08010-002)Program for New Century Excellent Talents in University of Ministry of Education of China(NO.NCET-08-0693)+1 种基金Fund for Jilin Province Excellent Innovation Team(NO.20111815)Project from Technology Bureau of Changchun City(NO.09YJ24)
文摘[ Objective ] This study aimed to investigate the expression of IPT gene directed by GmSARK promoter in Arabidopsis. ~ Method J IPT gene and C.m- SARK promoter were respectively cloned to construct plant expression vector and transform Arabidopsis. Tl transgenie plant lines were detected by PPT （ phosphino- thricin） herbicide selection and treated under drought and dark conditions for semi-quantitive RT-PCR analysis. E Result] GmSARK： ：IPT fusion gene was success- fully cloned and the plant expression vector p3301-GmSARK-IlYF was constructed. RT-PCR analysis showed that the target gene was expressed in T1 transgeuie plant lines at the mRNA level. [ Conclusion] This study laid the foundation for further investigating the roles of IPT gene directed by GmSARK promoter in plant stress resistance.
基金This work was supported by the Foundation for Ph. D. Student Research Station of the Ministry of Education.
文摘Fruit specific promoter (2A12) from Lycopersi-com esculentum and cDNA of isopentenyl-transferase (ipt) from Ti plasmid of Agrobacterium tumerfaciens C58 were cloned by PCR procedure respectively. Two plant expression vectors with 2A12/gus or 2A12/ipt were respectively constructed. These two chimeric genes were transferred into tomato by Agrobacterium mediated procedure. The results of Southern hybridization showed that the fusion genes had been integrated into tomatoes. The result of gus histochemi-cal staining showed that 2A12 had high fruit specific expressive capability in transgenic tomato. The ipt expression resulted in accumulation of high level of cytokinins (CTKs) in fruit lead to developmental changes in fruits and seeds. The fruit of ipt transformed tomato had the hyperplastic placenta with very few seeds or even seedless. The shelf life of transgenic fruits elongated for 1-2 weeks. The ratio of fruit set, the dry weight of fruit and the crude protein content in fruit were increased, while
文摘The isopenteryl transferase ( ipt ) gene from Agrobacterium tumefaciens (Smith et Townsend) Conn was driven under the tobacco TA29 promoter and introduced into tobacco (Nicotiana tabacum L.) plants by A. tumefaciens mediated transformation. PCR and Southern blot analysis confirmed that the ipt gene has integrated into the genomes of tobacco plants. The expression pattern of this chimeric TA29_ ipt gene in the transgenic plants was studied, and the endogenous cytokinin level in different organs was assayed by ELISA method. The results showed that the cytokinin content in the androecium of transgenic plants increased 3-4 times as compared with the control, and some changes of the fertility of the TA29_ ipt transgenic plants have been observed.
