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Plant expression carrier for improving plant cuttage taking root and apical dominance and its use

A plant expression vector and apical dominance technology, applied in the field of DNA recombinant vectors, can solve the problems of weakened apical dominance, shortened internodes, phenotypic variation, etc., and achieve the effect of promoting apical dominance and high rooting ability

Inactive Publication Date: 2005-10-19
陈晓阳 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the rolB gene is the most important root-inducing factor) to promote the rooting ability of plants, but some transgenic plants also produce phenotypic variations, such as weakened apical dominance, shortened internodes, etc.

Method used

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  • Plant expression carrier for improving plant cuttage taking root and apical dominance and its use
  • Plant expression carrier for improving plant cuttage taking root and apical dominance and its use
  • Plant expression carrier for improving plant cuttage taking root and apical dominance and its use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] Example 1 Construction strategy of GH3 / rolB gene expression vector

[0015] The GH3 / rolB gene is cloned in the pUC19 plasmid vector, and contains an EcoR I restriction site at both ends of GH3 / rolB by sequencing, and an EcoR I restriction site in the middle of the β-galactosidase gene of the expression vector pBin19. Therefore, the EcoR I enzyme was selected, and the GH3 / rolB gene was inserted into the pBin19 plasmid by restriction restriction method. Screening of recombinant plasmids can use the blue-white method, PCR method and EcoR I digestion method.

Embodiment 2

[0016] Example 2 Construction strategy of 35S / GA20ox gene expression vector

[0017] The pttGA20ox gene was cloned in the pGEX-4T-2 plasmid. Considering that the expression of the pttGA20ox gene is driven by 35S, pBI121 was selected as the plasmid constructed by the plant expression vector. There are Xbal I, BamH I and Sac I at both ends of the GUS gene fragment Restriction sites are available. After the sequence analysis of the pttGA20ox gene, it was confirmed that the pttGA20ox gene was cloned by PCR method, and Xbal I and Sac I restriction sites were added to the 5' and 3' end primers, respectively. The size of the pttGA20ox gene fragment is about 1.55KB, so high-fidelity pfu was selected for PCR amplification.

Embodiment 3

[0018] Example 3 Strategies for the construction of bivalent gene expression vectors

[0019] On the basis of constructing the pBin19rolB and pBI121GA plant expression vectors, the bivalent plant expression vectors of these two genes were constructed. Through the analysis of restriction sites of pBin19rolB and pBI121GA plasmids, the Kpn I and Sal I sites in pBin19rolB are suitable for constructing bivalent vectors, but there is no such site in pBI121GA plasmid, so PCR method was chosen for construction. The size of the entire 35S / GA / nos is about 2.6KB, and pfu cannot amplify fragments larger than 2.0KB, so another high-fidelity DNA polymerase, PyrobestTM DNA polymerase, was selected. This DNA polymerase has the same fidelity as pfu, and has the same amplification efficiency as Taq DNA polymerase, and can amplify 8KBp DNA fragments very well

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Abstract

The present invention relates to a kind one plant expression vector and its application. The plant expression vector is constituted via inserting plasmid pBin19 into two genes pttGA20ox and rolB, which are driven with 35S and GH3 promoter separately. Transforming the vector into plant can obtain transformed plant with greatly raised cutting rooting capacity, obviously strengthened apical dominance and no obvious morphological variation. Therefore, the plant expression vector the present invention constitutes may be used in transforming tree variety with hard cutting rooting to raise its cutting rooting capacity and strengthen its apical dominance.

Description

【Technical field】 [0001] The invention relates to a DNA recombination vector, in particular to a bivalent gene plant expression vector capable of promoting plant cutting rooting and top dominance and application thereof. 【Background technique】 [0002] A lot of researches have been done at home and abroad on improving the rooting ability of plant cuttings by using genetic engineering technology. Wherein, the rolB gene in the Agrobacterium rhizogenes (Agrobacterium rhizogenes) Ri (Root inducing) plasmid is used (there are four rol gene loci distributed in the TL-DNA region of the agrobacterium rhizogenes Ri (Root inducing) plasmid, named respectively as rolA gene and rolB gene , rolC gene and rolD gene. Among them, rolB gene is the most important root-inducing factor) to promote plant rooting ability to study the most, but some transgenic plants also produce phenotypic variation, such as weakened apical dominance, shortened internodes, etc. The results of hormone content det...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82
Inventor 陈晓阳李伟
Owner 陈晓阳
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