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Transgenic rice culture method

A transgenic, rice technology, applied in horticultural methods, botanical equipment and methods, cells modified by the introduction of foreign genetic material, etc., can solve problems such as randomness and uncertainty

Inactive Publication Date: 2010-01-20
ZHEJIANG UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, which site of the exogenous Bt gene integration in the rice genome is random and uncertain

Method used

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  • Transgenic rice culture method
  • Transgenic rice culture method
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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1: Identification of Specific Sites

[0048] The specific site of the present invention is analyzed and identified through the following test materials and test procedures.

[0049] (1) Plasmid vectors containing foreign genes

[0050] In the present invention, two plasmid DNAs are used for rice transformation (see figure 1 ). Among them, the plasmid pFHBT1 carries the Bt insect resistance gene coding sequence fused by cry1Ab / cry1Ac, and is driven by the ActinI promoter and nos terminator that can be effectively expressed in the genome of grass crops. The plasmid was constructed and used by academician Fan Yunliu of the Biotechnology Research and Development Center of the Chinese Academy of Agricultural Sciences and others. Plasmid pGL2RC7 carries hygromycin-resistant gene and anti-fungal chitinase gene coding sequence, both of which are expressed by the tobacco mosaic virus promoter CaMV35S and terminator nos. In the present invention, the plasmid is used f...

Embodiment 2

[0099] Example 2: Isolation, sequencing analysis and chromosome mapping of composite transgene constructs

[0100] The composite transgene structure includes the entire primary DNA structure of the foreign insertion sequence and the sequence flanking the integration site. To clarify the compound transgene structure, it is first necessary to isolate the flanking sequence adjacent to the insertion site, and then isolate the exogenous DNA sequence on the insertion site. After sequencing, the entire DNA primary structure can be determined. The chromosomal location of the insertion site can be determined by comparing and analyzing online bioinformatics using the known genome sequence of rice.

[0101] The isolation of the flanking sequences adjacent to the insertion site was carried out using TAIL-PCR (Thermal asymmetric interlaced-PCR) invented by Liu Yaoguang et al. TAIL-PCR uses a set of nested specific primers and short random degenerate primers (Shortarbitary degenerate primers...

Embodiment 3

[0133] Example 3: Sexual hybridization of exogenous Bt transgene composite structure

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Abstract

The provided specific site can integrates exogenous gene to stable express and affect little to the acceptor. Besides, it also provides a composite gene structure, which can be integrated by rice gene DNA sequence near specific site and DNA sequence of exogenous gene, and be used to culture new rice series by sexual hybridization and somatic hybridization.

Description

technical field [0001] The invention relates to the technical field of rice molecular breeding, in particular, the invention relates to a method for cultivating transgenic rice using a specific site on the rice genome and the obtained transgenic rice cells. technical background [0002] One of the main aspects of improving rice traits is to improve its insect resistance. Lepidoptera pests such as rice stem borer are one of the main limiting factors for high and stable rice yields. So far, there are no natural rice seed resources in China or even in the world that are resistant to Lepidoptera pests such as rice borer. In the past, the control of rice borer mainly relied on the use of chemical pesticides, but the use of a large number of pesticides not only caused serious residual pollution to the biological environment, but also reduced the hygienic quality of rice, thus affecting people's health. In addition, since borer larvae only stay on the outer surface of rice plants ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/10C12N15/82A01H4/00
Inventor 凃巨民张启发黄海清潘刚何予卿张集文
Owner ZHEJIANG UNIV
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