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Paulownia UDP-glucose pyrophosphorylase gene and use thereof

A technology of tunguridine diphosphate and uridine diphosphate, which is applied in the fields of application, genetic engineering, plant gene improvement, etc., can solve the problems of limited application range and application value, low strength and hardness, low bulk density, etc., and achieve improved plant The effect of quality or material

Inactive Publication Date: 2009-05-20
河南省绿士达林业新技术研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] But it should be noted that the other side of paulownia wood material: light (low bulk density), low strength and hardness, which seriously limits its application range and application value in construction, transportation, home decoration, furniture, papermaking, etc.

Method used

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  • Paulownia UDP-glucose pyrophosphorylase gene and use thereof
  • Paulownia UDP-glucose pyrophosphorylase gene and use thereof
  • Paulownia UDP-glucose pyrophosphorylase gene and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Cloning and sequencing of Paulownia partial UGP coding sequence

[0024] 1. Design of degenerate primers: the homology of plant UGP amino acids was analyzed by GenBanK BLAST system, and its complete homology part was selected, and degenerate primers were selected. All PCR primers involved in the present invention are listed below.

[0025] (1) Degenerate primers:

[0026] BUF 5'AARGAYGGNTGGTAYCCNCCNGG 3'

[0027] BUR 5'GGRTTNGGDATDATYTCCATYTT 3'

[0028] (2) 5' RACE primer:

[0029]Outer Primer 5'-taccgtcgttccactagtgattt-3'

[0030] Inner Primer 5'-cgcggatcctccactagtgatttcactatagg-3'

[0031] U.5GSP1 5’TGATTGCCTGTAAGTTGACCC 3’

[0032] U.5GSP2 5’GACATATTCTTTGCCCTGCGATA 3’

[0033] (3) 3'RACE Primer 3'RACE:

[0034] Outer Primer 5'-taccgtcgttccactagtgattt-3'

[0035] Inner Primer 5'-cgcggatcctccactagtgatttcactatagg-3'

[0036] U.3GSP1 5’TTGTTATCGCAGGGCAAAG 3’

[0037] U.3GSP2 5’ACACTAGCTGATGTGAAAGGTGGT 3’’

[0038] (4) Antisense gene primers:

[00...

Embodiment 2

[0070] Embodiment 2: Obtaining of unknown sequence upstream of Paulownia UGPmRNA 5':

[0071] A preferred solution is to use the 5'-FULL RACE kit (Code No: D315) produced by TaKaRa Company. The first step of this technical system is phosphorylation, the second step is to remove the cap, and the third step is to add a linker. Theoretically speaking, the first step eliminates all those that have lost their "caps", broken in the middle, and incomplete. Incomplete and incomplete mRNA, so that the unknown 5' end of the clone is complete, and the cloned mRNA may be full-length. At the same time, technologies such as adding specific adapters and nested PCR are used to amplify the full-length sequence of the 5' end of cDNA with high sensitivity and high specificity.

[0072] The partial coding sequence of Paulownia UGP was obtained by RT-PCR based on the obtained degenerate primers, and 2 specific nested PCR primers were optimized [see 1 in Example 1 (design of degenerate primers)]. ...

Embodiment 3

[0075] Embodiment 3: Obtaining of unknown sequence downstream of Paulownia UGP mRNA 3':

[0076] A preferred solution is to use 3'-FULL RACE Core Set Ver.20 (Code No: D314) produced by TaKaRa Company. This technology system uses technologies such as Oligad (T) adapters and primers added with specific sequences, so that the 3' end sequence of cDNA can be amplified more sensitively and specifically through the known partial cDNA sequence.

[0077] The partial coding sequence of Paulownia UGP was obtained by RT-PCR using the obtained degenerate primers, and 2 specific nested PCR primers were optimized [see 1 in Example 1 (design of degenerate primers)].

[0078] Concrete cloning procedures include: a. Extraction of total RNA of Paulownia: same as 2 (extraction of total RNA of Paulownia) in Example 1; b. Nested PCR reaction: the specific primer U.3GSP1 and 3' RACE OuterPrimer tested outside first [See 1 in Example 1 (design of degenerate primers)], perform PCR reaction, and then ...

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PUM

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Abstract

The invention discloses a paulownia uridine diphosphate glucose pyrophosphorylase gene and application thereof. Amino acid residue sequence of the paulownia uridine diphosphate glucose pyrophosphorylase gene is shown in SEQ ID No. of 1. The information can be used for constructing the paulownia uridine diphosphate glucose pyrophosphorylase gene, or antisense gene and RNA disturbing mass of the gene, which can be used for improving plant materials and constructing new materials with higher or lower cellulose content.

Description

Technical field: [0001] The invention discloses a uridine diphosphate glucose pyrophosphorylase (UDP-glucosepyrophosphorylase UGP) gene and its clone, belonging to the field of bioengineering, in particular to Paulownia. (Seem) Hemsl]) encoding 4CL gene and its application. Background technique: [0002] 1. Regarding the material improvement of Paulownia and its wood: [0003] Paulownia (Paulownia) is a deciduous tree of the Paulowniaceae family (Paulowniaceae), native to my country, except for a few species [P. fortunei (Seem) Hemsl.] distributed to Vietnam and Laos, other species are unique to our country. [0004] Paulownia is one of the three high-quality, fast-growing timber tree species in the world. It is widely distributed in 23 provinces, municipalities and autonomous regions. In many areas, it is used as the main tree species for tree planting, agricultural paulownia intercropping, and forest network construction; some places are gradually developing into mountai...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/14C12N15/55C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21A01H1/00
Inventor 陈占宽杨艳坤熊治国李荣幸穆守义叶金山王军军苗丽娟
Owner 河南省绿士达林业新技术研究所
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