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Sindora glabra sesquiterpenoid synthase SgSTPS2 as well as encoding gene and application thereof

A technology of sesquiterpene synthase and coding gene, which is applied in application, genetic engineering, plant genetic improvement, etc.

Active Publication Date: 2018-12-28
RES INST OF TROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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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  • Sindora glabra sesquiterpenoid synthase SgSTPS2 as well as encoding gene and application thereof
  • Sindora glabra sesquiterpenoid synthase SgSTPS2 as well as encoding gene and application thereof
  • Sindora glabra sesquiterpenoid synthase SgSTPS2 as well as encoding gene and application thereof

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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 sindora glabra sesquiterpenoid synthase SgSTPS2 as well as an encoding gene and an application thereof. The novel sesquiterpenoid synthase, namely, sindora glabra sesquiterpenoid synthase SgSTPS2 as well as the encoding gene thereof are provided. The protein can generate active sesquiterpenoid synthase after prokaryotic expression, catalyzes FPP (farnesyl pyrophosphate) orGPP (geranyl pyrophosphate) to generate multiple sesquiterpene or monoterpene compounds and can be used for 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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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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