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Site-specific integration transgenosis method for Arabidopsis thaliana L.

A fixed-point integration and transgenic technology, which is applied in the field of fixed-point integration and transgenesis of Arabidopsis thaliana, can solve the problems of long experimental period and low transformation efficiency, and achieve the effect of short experimental period, high transformation efficiency and great practical application value

Inactive Publication Date: 2010-11-24
INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
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AI Technical Summary

Benefits of technology

This patented technology allows for specific insertion into different parts or sections of DNA within certain genes that can be used to modify their expression levels without affecting other important aspects such as cell growth rate. It also simplifies the process of creating new crops from existing crop material. Overall this innovation makes it possible to improve agricultural production while reducing costs associated therewith.

Problems solved by technology

This patented technical problem addressed in this patents relates to improving the efficient use of introduced genes for targeting specific modifications within crops without causing damage from environmental factors like temperature fluctuations (temperature) and pH changes. Current methods involve either physically removing existing ones through excision or other processes, leading to reduced efficacy and potential harmful side effects.

Method used

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  • Site-specific integration transgenosis method for Arabidopsis thaliana L.
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  • Site-specific integration transgenosis method for Arabidopsis thaliana L.

Examples

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Embodiment Construction

[0047] The methods used in the following examples are conventional methods unless otherwise specified. The primers were all synthesized by IDT Company, the PCR kit was from Takara Company, the plant genomic DNA extraction kit (DNeasy miniprepkit) was purchased from QIAGEN Company, the nested PCR kit (Vectorette II Kit) was purchased from Sigma-Genosys Company, and the plant total RNA was extracted Kit (The Spectrum TM Plant Total RNA Kit) was purchased from Sigma-ALDRICH company, real-time quantitative PCR kit (EXPRESS Two-Step GreenER TM with Premixed ROX) was purchased from Invitrogen, and Basta reagent was purchased from AgroEvoUSA. Routine reagents were purchased from Takara Company. The strains used in the experiment are commonly used strains.

[0048] (1) Preparation of Agrobacterium strains containing target sites

[0049] The Agrobacterium strain is GV3101, and the expression vector contains a promoter-loxP-Cre coding frame structure and a bar selection marker ...

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Abstract

The invention discloses a site-specific integration transgenosis method for Arabidopsis thaliana L., which comprises the following steps of: integrating target sites into genomes of target plants with agrobacterium rhizogenes by utilizing a florescence spraying method, and screening single-copy heterozygote transgenic plants out; and integrating target genes into the genomes of the screened single-copy heterozygote transgenic plants with the agrobacterium rhizogenes by utilizing the florescence spraying method again, and screening specific-site integrated homozygote plants out as successfully specific-site integrated plants. The site-specific integration transgenosis method for the Arabidopsis thaliana L. has the advantages of high conversion rate and short cycle.

Description

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Claims

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Application Information

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Owner INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
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