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A kind of Phoebe sesquiterpene synthase sgstps2 and its coding gene and application

A technology of sesquiterpene synthase and eucalyptus, applied in application, genetic engineering, plant genetic improvement and other directions

Active Publication Date: 2021-04-27
RES INST OF TROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the biosynthetic genes for terpene synthases in Phoebe oleifera have not been isolated and identified

Method used

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  • A kind of Phoebe sesquiterpene synthase sgstps2 and its coding gene and application
  • A kind of Phoebe sesquiterpene synthase sgstps2 and its coding gene and application
  • A kind of Phoebe sesquiterpene synthase sgstps2 and its coding gene and application

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Experimental program
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Effect test

Embodiment 1

[0028] 1. Clone the oleifera sesquiterpene synthase gene SgSTPS2, construct a cloning vector and transform prokaryotic cells

[0029] Extract the RNA of the stem tissue of Phoebe oleifera, and use the reverse transcriptase M-MLV to reverse transcribe the synthesized cDNA. Using the cDNA as a template, the forward primer is: 5'-ATGTCGGTTGCAGCTTTAGC-3', the reverse primer is: 5'-TCATGCGACAGGATTTATGA-3', and PCR amplification is performed using Ex Taq DNA polymerase from TakaRa Company; the PCR conditions are: : 94°C for 5min; 94°C for 30s, 55°C for 1min, 72°C for 1min30s, 35 cycles; 72°C for 10min. The PCR product was detected by 1% agarose gel electrophoresis, and the results were as follows: figure 1 as shown, figure 1 M is the DNA marker DL2000, and the fragment size of the target gene SgSTPS2 is about 1700bp, which is in line with expectations.

[0030] Use the method of agarose gel electrophoresis gel recovery kit to recover the target gene fragment, carry out TA cloning...

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Abstract

The invention discloses a oleifera sesquiterpene synthase SgSTPS2 and its coding gene and application. The present invention provides a new sesquiterpene synthase, SgSTPS2 and its coding gene, the protein can generate active sesquiterpene synthase after prokaryotic expression, and catalyze farnesyl pyrophosphate (FPP) or geranyl pyrophosphate (GPP) to generate a variety of sesquiterpenoids or monoterpenoids, which can be used for their mass production.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, and in particular relates to an oleaginous sesquiterpene synthase SgSTPS2 and its coding gene and application. Background technique [0002] Sindora glabra Merr.ex de Wit belongs to the genus Sindora glabra Merr.ex de Wit. It is originally distributed in Hainan Province of my country. It is a tall tree in the tropics and subtropics. The most important feature of Phoebe oleifera is that resin oil can be secreted after the xylem of the trunk is damaged. Its flammability is similar to diesel oil, so it is also called "diesel tree". It has a wide range of uses in oil, medicine, ornamental and wood. development and utilization prospects. [0003] The components of oleoresin oil mainly include sesquiterpenoids, accounting for more than 70%, and diterpenoids, accounting for more than 14%. Phoebe oil also contains active ingredients α-amorphene, cubeba oleene, α-humulene, cycloalfalen...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/88C12N15/60C12N15/70C12N1/21C12P5/00C12P7/04C12P15/00C12R1/19
CPCC12N9/88C12P5/002C12P5/005C12P7/04C12P15/00
Inventor 余纽杨锦昌李荣生尹光天邹文涛何畅
Owner RES INST OF TROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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