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Peanut diacylglycerol acyltransferase (DGAT2), and coding gene and application thereof

A technology of diacylglycerol acyl transferase, applied in the field of molecular biology and biology, can solve the problem of lack of DGAT1 and DGAT2

Inactive Publication Date: 2011-10-19
山东省农业科学院高新技术研究中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, so far, there is no information about DGAT1 and DGAT2 in peanut

Method used

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  • Peanut diacylglycerol acyltransferase (DGAT2), and coding gene and application thereof
  • Peanut diacylglycerol acyltransferase (DGAT2), and coding gene and application thereof
  • Peanut diacylglycerol acyltransferase (DGAT2), and coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Cloning of rate-limiting enzyme gene AhDGAT2a of triglyceride synthesis pathway in peanut

[0041] (1) RNA extraction: The CTAB method was used to extract the total RNA of immature seeds of peanut variety Luhua 14 (Arachis hypogaea L.), and the specific steps were as follows:

[0042] a) Weigh 0.2g of peanut material or a sample stored at -70°C, fully grind it into a powder in liquid nitrogen, and add liquid nitrogen slowly during the grinding to prevent the material from thawing;

[0043] b) Quickly transfer the ground tissue to a preheated (65°C) EP tube containing 600 μl CTAB extract, and immediately vortex vigorously for 30 seconds to make it evenly mixed;

[0044] c) Water bath at 65°C for 2-5 minutes, during which time vortex vigorously for 5 times;

[0045] d) After cooling, add an equal volume of chloroform / isoamyl alcohol, vortex for 1 min, mix well, and centrifuge at 12,000 rpm for 15 min at 4°C;

[0046] e) Transfer the supernatant to another new EP tube, a...

Embodiment 2

[0107] As described in Example 1, the difference is that during the construction of the peanut AhDGAT2a gene plant expression vector, the constructed plant overexpression vector was named pROK II-35S-AhDGAT2b.

Embodiment 3

[0109] As described in Example 1, the difference is that during the cloning process of the rate-limiting enzyme gene AhDGAT2c of the triglyceride synthesis pathway in peanut, the peanut variety used is Feilongxiang (Arachis hypogaea L.).

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Abstract

The invention relates to a peanut diacylglycerol acyltransferase (DGAT2), and a coding gene and application thereof, belonging to the technical fields of molecular biology and biotechnology. The full-length sequence of the DGAT2 gene is cloned from peanut for the first time, and is named AhDGAT2 (Arachis hypogaea DGAT). Eight homologous genes of DGAT2 are obtained from different peanut species, and are respectively named AhDGAT2a, AhDGAT2b, AhDGAT2c, AhDGATT2d, AhDGAT2e, AhDGAT2f, AhDGAT2g and AhDGAT2h. The gene has difference of specific amino acids on the N-terminal of the coding protein, but is identical at the C-terminal. The gene can be used for transgenic breeding of oil crops to increase the oil content in the oil crops.

Description

technical field [0001] The invention relates to peanut diacylglycerol acyltransferase DGAT2 and its coding gene and application, belonging to the technical fields of molecular biology and biotechnology. Background technique [0002] Peanut is one of the important oil crops and economic crops in my country, with a planting area of ​​about 5.025 million hectares, and its total output ranks first in the world. Peanut contains about 50% fat. Among several major edible oil crops, its fat content is second only to sesame, but higher than rapeseed, soybean and cottonseed. Vegetable oil (triacylglycerol) is one of the most abundant and important organic compounds in oil plants. It not only plays an important role in the process of plant growth, development and reproduction, but also as a renewable bioenergy product, has very Wide range of applications. Vegetable oil is the main source of edible lipids, accounting for about 75% of the world's lipid consumption, and many monounsatur...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/54C12N9/10C12N15/63C12N1/21C12N15/82A01H5/00
Inventor 毕玉平彭振英王龙龙纪鸿飞李兰陈高单雷
Owner 山东省农业科学院高新技术研究中心
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