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Plant polygene expression vector composition and application thereof

An expression vector and multi-gene technology, applied in plant gene improvement, application, genetic engineering, etc., can solve the problem of rarely allowing more than one target gene to be cloned and transferred, and achieve stable expression, increased transformation efficiency, and increased stability Effect

Inactive Publication Date: 2011-05-25
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although these vectors have been proven to be operable in the field of botany research and biotechnology, the basic design of these vectors is very restrictive and rarely allows the cloning and transfer of more than one gene of interest (except for screening genes)

Method used

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  • Plant polygene expression vector composition and application thereof
  • Plant polygene expression vector composition and application thereof
  • Plant polygene expression vector composition and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0041] Embodiment 1, the construction of plant polygene expression vector combination

[0042] The present invention considers that instead of assembling the gene or DNA fragment into the final vector with a large scale (about 10kb), it is better to first ligate the gene or DNA fragment to two intermediate vectors with smaller scales, The two DNA fragments (intermediate vector) are then assembled into a complete final expression vector using recombinase-mediated site-specific recombination. In this way, even the same set of multiple cloning sites can be used separately on the intermediate vector, and the plant selection marker gene of the final expression vector can also be conveniently and quickly transformed by using the intermediate vector. Therefore, the plant polygene expression vector combination of the present invention is composed of two expression vectors, vector A and vector B, and the specific features are:

[0043] Both have a specific recombination site (eg Loxp)...

Embodiment 2

[0071] Embodiment 2. Effect verification of the combination of plant polygene expression vectors of the present invention

[0072] The plant multigene expression vector combination vector A and vector B prepared in Example 1 can be respectively added to the exogenous gene expression cassettes that need to be transferred into plants, and then recombined and integrated into a plant expression vector that can express multiple genes. Among them, pModularB-gfp already carries an exogenous gene GFP.

[0073] Taking the following method as an example, the effect of the combination of the plant polygene expression vector of the present invention will be described.

[0074] 1. Insert foreign gene expression cassette into vector A

[0075] 1) Obtaining the recombinant vector containing betaine aldehyde dehydrogenase (BADH) gene expression cassette

[0076] According to GENEBANK No. DQ497233, the betaine aldehyde dehydrogenase (BADH) gene in the plant spinach was artificially synthesiz...

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Abstract

The invention discloses a plant polygene expression vector composition and application thereof. The vector composition consists of a vector A and a vector B; the nucleotide sequence of the vector A comprises a T-DNA terminal sequence, a multiple cloning site sequence and a specific recombinant site sequence sequentially connected in series; and the nucleotide sequence of the vector B comprises a specific recombinant site sequence, a multiple cloning site sequence and a T-DNA terminal sequence sequentially connected in series. The plant polygene expression vector composition is reduced to minimum, and copy number, stability and conversion efficiency of plasmids are improved. Under the condition of ensuring that the conversion efficiency is not remarkably reduced, the smaller the plasmids are, the longer the fragment of the containable exogenous DNA is, so the vector composition is very favorable for experiment operation.

Description

technical field [0001] The present invention relates to a combination of plant polygene expression vectors and its application. Background technique [0002] In the past few decades, scientists have developed a large number of vectors suitable for cloning, transferring and expressing foreign genes in plant cells. Although these vectors have proven operable in the fields of botanical research and biotechnology, the basic designs of these vectors are very restrictive and rarely allow cloning and transfer of more than one gene of interest (except for screening genes). The growing focus on dissecting complex metabolic pathways and uncovering the potential of polygenic traits for plant bioengineering has led to the need for new methods and tools to transform multiple genes simultaneously. [0003] As the main method of plant genetic engineering, Agrobacterium vectors have been continuously improved and modified with different research purposes in the past few decades. Throughou...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/84
Inventor 王涛马磊董江丽
Owner CHINA AGRI UNIV
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