Method for obtaining seed-free plant by utilizing genetic engineering technology and application thereof

A genetically engineered, seedless technology that can be used in the field of genetic engineering to solve problems such as toxicity, strong cells, and host cell death

Inactive Publication Date: 2012-02-15
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As a small molecule RNase, the expression of Barnase in cells can lead to non-specific degradation of intracellular RNA, which manifests as strong cytotoxicity, and its expression in specific cells usually leads to the death of host cells

Method used

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  • Method for obtaining seed-free plant by utilizing genetic engineering technology and application thereof
  • Method for obtaining seed-free plant by utilizing genetic engineering technology and application thereof
  • Method for obtaining seed-free plant by utilizing genetic engineering technology and application thereof

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Embodiment Construction

[0037] Below in conjunction with accompanying drawing and embodiment describe in detail:

[0038] The present invention intends to adopt the transgenic method mediated by Agrobacterium to achieve the expected goal. The specific method is as follows:

[0039] 1. Gene fragment cloning

[0040] (1) Cloning of NapinB (a major storage protein in rapeseed) gene promoter sequence

[0041] According to the upstream untranslated region sequence of NapinB gene (Genbank accession number: X14492), PCR primers P1: TCC TGC GTT ATC CCC TGA TTC TG and P2: ATA ACT TCG TAT AAT GTATGC TAT ACG AAC GGT ATC GTG TAT GTT TTT AAT were designed and synthesized CTT GTT TG. Using the genomic DNA of rapeseed Zhongyou 821 as a template, the promoter sequence of NapinB gene was cloned. The amplified sequence is called Fragment 1.

[0042] (2) Barnase (ribonuclease) gene fragment cloning

[0043] Because the leaky expression of Barnase in prokaryotes will cause the death of host cells, it cannot be dir...

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Abstract

The invention discloses a method for obtaining seed-free plant by utilizing a genetic engineering technology and application thereof, relating to the technical field of genetic engineering. The method comprises the following steps: (1) cloning or synthesizing a seed-specific expressed rape NapinB gene promoter; (2) cloning or synthesizing a Barnase gene segment; (3) cloning or synthesizing an Actin gene intron of a host plant; (4) constructing a genetic transformation vector with a Barnase gene containing the intron as a target sequence; and (5) transforming and screening a transgenic plant. By using the method, a seed plant can be transformed to obtain the seed-free character; as a parent, the obtained transgenic variety can be hybridized with any parent so as to obtain a seed-free hydrid; and the obtained transgenic variety dose not influence the seed-free character when foreign pollen performs pollen changing.

Description

technical field [0001] The present invention relates to the technical field of genetic engineering, in particular to a method for obtaining seedless plants using genetic engineering technology and its application; specifically, the present invention is controlled by the NapinB (a main storage protein in rapeseed seeds) gene promoter Barnase (ribonuclease) gene expression, constructs the method for vector transformation plant to obtain seedless character; Its application is to produce seedless plant based on this method and keep the seedless characteristic of plant from changing because of pollination hybridization. Background technique [0002] Seedless breeding is an important direction of melon and fruit breeding, and many seedless fruits have been put on the market now, such as seedless watermelons, seedless grapes, and seedless kiwis. At present, most of the seedless fruits on the market are obtained by breeding triploids or inducing parthenocarpy. [0003] In recent ye...

Claims

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Application Information

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IPC IPC(8): C12N15/63A01H5/00A01H1/02
Inventor 卢长明陈桂敏吴刚武玉花草应龙
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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