Production process of transgenic rape as sinapic acid bioreactor

A bioreactor, transgenic technology, applied in the fields of biochemical equipment and methods, plant genetic improvement, and botany equipment and methods

Inactive Publication Date: 2003-08-20
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] Although there have been many reports on the use of genetic engineering technology to breed super-high mustard rapeseed, the erucic acid content of the obtained transgenic rapeseed is also below 60%.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0038] The following examples illustrate the implementation of the present invention in detail, but the present invention is not limited to the application provided below, and the following description does not constitute any sense of limitation to the protection scope of the present invention.

[0039] Material:

[0040] Plant material: Brassicanapus varieties with relatively high oil and erucic acid content were used.

[0041] Strains and plasmids: E. coli HB101, E. coli JM109, A. tumefaciens EHA101, plasmids pBS SK+, pUC19, super binary vectors pIG121 and pBI121.

[0042] Enzymes and chemical reagents: restriction endonuclease, calf intestinal alkaline phosphatase, T4-DNA ligase, TaqDNA polymerase were purchased from MBI Company, DNA molecular weight marker λDNA / HindIII, PCR Marker were purchased from Huamei Biotechnology Company, PCR Primers were synthesized by Sangon Biotechnology Company, X-gal, IPTG, and X-glue were purchased from Sigma Company, random primer label...

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Abstract

The production process of transgenic rape as sinapic acid bioreactor includes the steps of: cloning phosphoenolpyruvate carboxyilase gene PEP segment and constituting it into Anti-PEG gene plant expression vector, introducing Anti-PEP gene into rape genome through gene conversion process to obtain "high-oil rape" with efficient oil synthesizing rape seed; preparing lytic phosphatidic acid acyltransferase (LPAAT) gene capable of connecting sinapic acid to the sn-2 site of 3-glycerol phosphate and connecting it to plant expression vector promoter to constitute LPAAT gene plant expression vector; introducing LPAAT gene into rape genome to obtain "high-sinapic acid rape" with efficient sinapic acid and 3-glycerol phosphate combining seed; crossbreeding "high-oil rape" and "high-sinapic acid rape" to obtain F1 generation rape plant with both "high-oil rape" characteristic and "high-sinapic acid rape" characteristic.

Description

technical field [0001] The invention relates to plant breeding, in particular to changing the fatty acid synthesis pathway of rapeseed by using genetic engineering technology to use rapeseed as a bioreactor for erucic acid. technical background [0002] In addition to edible, nearly one-third of the vegetable oil produced in the world is used in non-edible products. It is very simple to separate and extract various oil compounds from oil plant seeds, avoiding the production of harmful substances to the environment [1，2] . [0003] Erucic acid is an important chemical raw material. Rapeseed seeds contain about 40% (w / w) oil, the highest of all oilseeds [3] , wherein the theoretical maximum value of erucic acid content accounts for 66.6% (w / w) of oil. Using rapeseed as erucic acid bioreactor to cultivate high erucic acid special industrial rape varieties not only opens up a new field for agricultural production, but also provides cheap and renewable ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H1/00A01H1/02A01H5/00C12N15/54C12N15/60C12N15/63
Inventor 陈锦清黄锐之朗春秀刘智宏胡张华
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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