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Method for generating pterostilbene by utilizing grape resveratrol-oxygen-methyl transferase to catalyze resveratrol

A technology of methyltransferase and resveratrol, applied in the field of plant genetic engineering, can solve the problems that the biosynthetic pathway of pterostilbene has not been clearly studied

Inactive Publication Date: 2011-07-13
NORTHWEST A & F UNIV
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Problems solved by technology

[0005] Despite its high antifungal activity and its attractive pharmacological properties, the biosynthetic pathway of pterostilbene in grapevine has not been elucidated

Method used

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  • Method for generating pterostilbene by utilizing grape resveratrol-oxygen-methyl transferase to catalyze resveratrol

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Embodiment Construction

[0022] Below in conjunction with embodiment and accompanying drawing, the present invention is described in further detail:

[0023] a. Cloning of ROMT gene and construction of expression vector

[0024] Using the wild species of Vitis vitis Huadongensis strain "Baihe-35-1" as the material, according to the EST sequence of the ROMT gene of the Chinese wild grape Vitis huadongensis, ROMT was cloned by 5' and 3' RACE full-length gene cloning technology, and the gene open reading was obtained The frame length is 1074bp;

[0025] b. Construction of ROMT gene plant expression vector

[0026] b.1 Insert the ROMT open reading frame into the 35S promoter of the binary vector pWR306 to construct pWR306-ROMT,

[0027] b.2 Insert the open reading frame of the STS gene into the 35S promoter of the binary vector pWR306 to construct pWR306-STS as a positive control,

[0028] b.3 The empty vector pWR306 was used as a negative control,

[0029] b.4 Transform the constructed vectors into A...

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Abstract

The invention discloses a method for generating pterostilbene by utilizing grape resveratrol-oxygen-methyltransferase to catalyze resveratrol, grape ROMT (resveratrol-oxygen-methyl transferase) genes are cloned by utilizing 5' and 3' RACE (rapid amplification of cDNA ends) full-length gene cloning technology according to EST (expressed sequence tag) sequence of the ROMT genes of Chinese wild grapes in East China, and the full length of an open reading frame of the genes is 1074bp; and the cloned grape STS (steroid sulfatase) and ROMT genes can be simultaneously transferred into model plant tobacco, tobacco plant analysis is utilized for analyzing the biological synthesis path of generating the pterostilbene by utilizing the grape ROMT to catalyze the resveratrol, a grape ROMT gene sequence is obtained for orientation, and the ROMT genes can catalyze the resveratrol to generate the pterostilbene in the transgenic tobacco, thereby providing the sequence of the ROMT genes and the method for synthesizing the pterostilbene by utilizing plants and providing a basis for getting high-quality, disease-resistant and health care crops.

Description

technical field [0001] The invention relates to the construction of a recombinant expression vector derived from grape resveratrol-oxygen-methyltransferase gene and a method for synthesizing pterostilbene in transgenic tobacco, which belongs to the field of plant genetic engineering. Background technique [0002] The three phenolic hydroxyl groups of resveratrol have strong electron donating ability, which is the reason for the strong antioxidant capacity of resveratrol. The antioxidant activity of resveratrol hydroxyl analogs is related to the number and position of phenolic hydroxyl groups. Therefore, compounds obtained by increasing or decreasing the number of hydroxyl groups on the two benzene rings of resveratrol and changing the positions of hydroxyl groups often have the same pharmacological effects as resveratrol. Due to the three phenolic hydroxyl groups in the molecule, resveratrol is very unstable and easily oxidized to produce pterostilbene. Therefore, people tr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/09C12N15/82C12N15/54C12P7/22C12Q1/68
Inventor 徐炎王跃进许腾飞赵霄晨李凤菊李志谦
Owner NORTHWEST A & F UNIV
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