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Plant dualistic expression system for rejecting screening marked gene and application thereof

A technology for screening marker genes and expression systems, which is applied in the field of plant genetic engineering and can solve the problems of co-transformation of plants and low efficiency of screening marker genes

Active Publication Date: 2008-07-09
SINOBIOWAY BIO AGRI GRP CO LTD
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Problems solved by technology

However, these methods have the disadvantages of obtaining co-transformed plants and eliminating the selection marker gene with low efficiency.

Method used

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  • Plant dualistic expression system for rejecting screening marked gene and application thereof
  • Plant dualistic expression system for rejecting screening marked gene and application thereof
  • Plant dualistic expression system for rejecting screening marked gene and application thereof

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Embodiment Construction

[0033] The present invention will be further described below in conjunction with specific embodiments.

[0034] The methods used in the following examples are conventional methods unless otherwise specified. The primers used were synthesized by Beijing Saibaisheng Biotechnology Co., Ltd., and the sequencing was completed by Shanghai Boya Biotechnology Co., Ltd. The nucleic acid in the PCR kit and vector construction process The T4 DNA polymerase used for Dicer and blunt end was purchased from Zibao Bioengineering Co., Ltd., and the ligation kit was from Invitrogen Company. The methods were all carried out according to the methods provided in the kit. The vector pCAMBIA1301 used in the experiment was donated by the laboratory of Professor Wang Xiping of Fudan University; the vector pXQ-35S was the vector pCAMBIA2300-ACTIN (given by the laboratory of Professor Chu Chengcai, Institute of Genetics, Chinese Academy of Sciences) digested by BamHI and KpnI and linked to the vector pCA...

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Abstract

The invention discloses a plant two-element expression system which rejects selective marker genes and is composed of an objective gene expression vector and a selective marker gene vector, wherein the objective gene expression vector comprises a promoter, the downstream transcription activation factor gene and a multiple cloning site and the selective marker gene vector comprises a cis-acting element of the transcription activation factor and the downstream selective marker gene. The above vectors are used together to transform plant, the transcription activation factor is combined with the cis-acting element to activate the expression of the selective marker genes and achieve the aim of screening transgenic plant, the transcription activation factor separates from the acting element and the downstream selective marker gene due to the effect of meiotic division in future generations, and thereby transgenic plant stains which do not contain the selective marker genes are obtained. The plant transforming method for rejecting the selective marker genes has the advantages that the co-transformation efficiency is high, the selective marker genes can be ejected easily and conveniently, and the method can be used to foster transgenic plant with safety.

Description

technical field [0001] The invention relates to the field of plant genetic engineering, in particular to a binary expression vector system capable of removing selectable marker genes and its application in cultivating safe transgenic plants. Background technique [0002] Selectable marker genes play an important role in plant genetic engineering, and the use of selectable marker genes can screen transformed cells from a large number of untransformed cells in the process of plant transgenesis. However, the screening genes in transgenic plants have raised concerns about a series of biosafety aspects, such as whether the selectable marker genes encode toxic substances and allergens, whether it is not conducive to changes in plant metabolism, and whether it will reduce the efficacy of therapeutic drugs. , and whether it will migrate between closely related species and pathogens. How to remove selectable marker genes from transgenic plants has become a concern of people. At pre...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H1/00
Inventor 翟晨光张蕾夏勉
Owner SINOBIOWAY BIO AGRI GRP CO LTD
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