Superstrong fusion promoter, fusion method and use thereof

A fusion method and promoter technology, applied in the direction of using vectors to introduce foreign genetic material, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problem of low expression of foreign genes and achieve the effect of increasing production

Inactive Publication Date: 2003-03-12
SHANDONG AGRICULTURAL UNIVERSITY
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Problems solved by technology

[0003] Aiming at the problem of low expression of exogenous genes in transgenic technology, the present invention fuses octopine synthase (ocs) enhancer sequences and mannopine synthase (mas) promoter core sequences with different copy numbers to obtain A strong promoter for high-efficiency expression in plants, used to construct high-efficiency expression vectors

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  • Superstrong fusion promoter, fusion method and use thereof
  • Superstrong fusion promoter, fusion method and use thereof
  • Superstrong fusion promoter, fusion method and use thereof

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Embodiment 2

[0019]The mas promoter (-189 to +65) on the Agrobacterium Ti plasmid was amplified and cloned by PCR method, the mas promoter fragment in the cloning vector was digested with Hind III and Xba I, and the target fragment was recovered by electrophoresis. At the same time, the pBI121 plasmid was also digested with Hind III and Xba I, and the remaining vector fragment was recovered by electrophoresis. For the recovery method, refer to the instruction manual of the DNA gel recovery kit of Shanghai Sangon Biotechnology Co., Ltd. Ligate the remaining vector fragment after pBI121 plasmid digestion with the mas promoter fragment in the cloning vector with the recovered fragment after double digestion with Hind III and Xba I, transform Escherichia coli, and screen the recombinant through enzyme digestion identification, and the recombinant is named : pMAS. The ocs enhancer (-294 to -116) on the Agrobacterium Ti plasmid was amplified and cloned by PCR method, the ocs enhancer fragment i...

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Abstract

A ultrastrong fusion promoter oms4, its expression vector and its application in transgenic tobacco are disclosed. Said oms 4 promoter is constructed with 4 copied ocs enhensers by cyclically forward linking them to the core fragment of mas promoter. The oms4 promoter can be used to replace the 35S promoter in expression vector pBI121 of plant for constructing pOMS4. Its advantage is high expression level of exogenous gene gus in tobacco (increased by 23-47 times).

Description

(1) Technical field [0001] The invention relates to a super-powerful promoter and its fusion method and application, belonging to the research field of eukaryotic gene expression regulation technology. Constructing the promoter into a plant expression vector can drive the high-efficiency expression of foreign genes in transgenic plants. (2) Background technology [0002] As an important element to initiate gene transcription, promoter has been studied in detail. People have discovered some strong promoters from different organisms, but with the deepening of genetic engineering, it is no longer enough to rely on these natural promoters to improve gene expression activity. Fusion promoters and binary promoters can efficiently drive the expression of exogenous genes, and have become a hotspot in promoter research. Fusion promoters connect the core promoter and enhancer sequences of heterologous promoters to construct super-transcriptional promoters. A dual promoter is a fusi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/62C12N15/67C12N15/82
Inventor 郑成超刘石娟杨国栋
Owner SHANDONG AGRICULTURAL UNIVERSITY
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