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Strawberry reverse transcription transposon gene and transcription characteristic

A technology of retrotransposon and gene characterization, applied in the direction of DNA / RNA fragments, recombinant DNA technology, etc., can solve problems such as the activation of retrotransposable elements

Inactive Publication Date: 2016-06-29
SHANDONG INST OF POMOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, retrotransposable elements may be activated as the number of passages of strawberry micropropagated plantlets increases

Method used

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  • Strawberry reverse transcription transposon gene and transcription characteristic
  • Strawberry reverse transcription transposon gene and transcription characteristic
  • Strawberry reverse transcription transposon gene and transcription characteristic

Examples

Experimental program
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Effect test

Embodiment 1

[0014] Example 1: Obtaining the full length of the FaREII gene.

[0015] In the early stage, we designed degenerate primers based on the conserved motifs of retrotransposon RT genes in other plants. Using PCR and RT-PCR techniques, we cloned two RT genes from strawberry cultivar 'All Star'. A Ty1-copia group retrotransposon FaREI was obtained by translocation technology, and it has been published on NCBI, and the Genbank accession number is FJ871121. In order to systematically study the relationship between retrotransposons and somaclonal variation in strawberry regeneration in vitro, we also studied the complete sequence of another retrotransposon. Chromosome walking was performed according to the principle and method of GenomeWalkerUniversalKit (Clontech), and the high-quality strawberry genomic DNA was digested with four kinds of restriction endonucleases DraI, StuI, EcoRV, PvuII, based on the strawberry retrotransposon RT gene sequence, Four genomic libraries for walking ...

Embodiment 2

[0017] Example 2: FaREII retrotransposon sequence analysis.

[0018] For the obtained FaREII retrotransposon, through homology search and multiple alignment for gene structure analysis and characteristic sequence query, it was found that the FaREII sequence has the gene structure characteristics of typical Ty1-copia retrotransposable elements, and the sequence The two ends of the LTR sequence are direct repeats, the PBS site is adjacent to the LTR sequence at the 5' end, the PPT site is upstream of the LTR at the 3' end, and the GAG-POL coding region is between the PBS and PPT sites. The encoded protein is according to The order of GAG-PR-INT-RT-RNaseH is arranged in a typical Ty1-copia class retrotransposon gene structure ( figure 2 ). According to the cluster analysis of FaREII and other Ty1-copia and Ty3-gypsy retrotransposable elements ( image 3 ), FaREII also clustered into the Ty1-copia class, again indicating that FaREII is a retrotransposon of the Ty1-copia class. ...

Embodiment 3

[0019] Example 3: Characteristic motif analysis of pol gene.

[0020] The pol gene of plant retrotransposons is not only the basic structural domain of retrotransposons, but also existing studies have shown that most of the pol genes of retrotransposable elements have their own conserved motifs, conserved Sequence variation can reflect the evolutionary relationship of retrotransposons. Contains the conserved sequence of the C-C-H-C nucleic acid binding domain in the GAG ​​region, and also contains the D(T / S)G characteristic sequence of the PR region; in the INT region, has the H-H-C-C motif of the zinc finger domain; in the RT region contains the motif YVDDML; RNaseH Regions of Motif I (KHI(F)D(E)) and Motif II (A(S)DX 2 TK). These conserved motifs are present in the pol gene of FaREII ( Figure 4 ).

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Abstract

The invention discloses a strawberry reverse transcription transposon gene sequence and a transcription characteristic analysis result. A full length of a gene DNA sequence is 5386 bp, and is named as FaREII, the gene sequence has a Ty1-copia reverse transcription transposon gene characteristic sequence, and is arranged according to gag and pol genes, wherein pol gene contains the characteristic motifs of GAG, PR, IN, RT and RH genes, a long terminal repeat sequence of 393 bp is positioned at two ends of the gene, and the 393 bp is a typical Ty1-copia reverse transcription transposon. According to the invention, the transcription of FaREII can be happened under hormone stress through Nothern Blot.

Description

technical field [0001] The invention relates to a retrotransposon gene, in particular to the retrotransposon gene derived from strawberry and the analysis of its transcriptional activity. Background technique [0002] Strawberry is a perennial evergreen herb that belongs to Rosaceae (Fragaria) in plant taxonomy. It is a small berry with high economic value. In recent years, domestic strawberry cultivation and production have developed rapidly. At present, the area and output of strawberries in China Both rank first in the world. Strawberry belongs to perennial herbaceous plant, adopts asexual reproduction in production. Long-term vegetative propagation resulted in the production and accumulation of more clonal variation in the main strawberry varieties. Although researchers have been able to breed bud varieties from the clonal mutant plants of strawberry (for example, 'Xinxing' was bred from 'All Star', and 'Kinugan' was bred from 'Nyofeng'), But mutations have also cause...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11
Inventor 何平李林光李慧峰王海波
Owner SHANDONG INST OF POMOLOGY
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