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A rice stamen-specific expression promoter osanth2 and its application

A promoter and stamen technology, which is applied in the field of biotechnology and crop genetic engineering, can solve the problems of limiting the ability to improve stamen traits, and the genetic resources of stamen-specific promoters are not very rich, so as to achieve the effect of improving rice fertility

Active Publication Date: 2020-04-21
RICE RES ISTITUTE ANHUI ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the genetic resources of stamen-specific promoters found so far are not very rich, which limits the improvement of people's ability to improve stamen traits

Method used

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  • A rice stamen-specific expression promoter osanth2 and its application
  • A rice stamen-specific expression promoter osanth2 and its application
  • A rice stamen-specific expression promoter osanth2 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Acquisition of OsAnth2 Promoter Containing Restriction Sites

[0035] Step 1. Design of primers

[0036] According to the whole genome sequence of rice variety Nipponbare (Oryza sativa L cv. Nipponbare) provided by NCBI, the amplification primers were designed according to the sequence of rice OsAnth2 gene, and the enzyme cutting sites of the primers were designed according to the characteristics of the selected vector and target gene.

[0037] In this embodiment, rice binary expression vector pCAMBIA1391( figure 1 In part A, from CAMBIA, openly used vectors, Anhui Academy of Agricultural Sciences, Ministry of Agriculture, Rice Group of Supervision, Inspection and Testing Center for GMO Product Components) as an example, the target gene is the Gus gene, and the specifically designed primers are: forward primer (SEQ ID No: 2) with HindIII at the 5' end, restriction site (AAGCTT), reverse primer (SEQ ID No: 3) with BamHI at the 5' end, restriction site (GGATCC), the prim...

Embodiment 2

[0054] In order to verify that the sequence in the sequence listing SEQ ID No.1 of the present invention still has a promoter function after removing the 22 bp at the front end and the 22 bp at the back end, the present invention also carried out experiments similar to the above Example 1 on this sequence using other primers , that is: use the rice variety Nipponbare DNA as a template, use new forward primers and reverse primers to amplify the promoter; recover the target fragment amplified by PCR, connect it to the PGEM-T-Easy vector, and transfer it to the activated sensory State cells; then, connect it with the pCAMBIA1391 fragment with T4 ligase to obtain the corresponding crop expression vector, and use the freeze-thaw method to transfer the crop expression vector into Agrobacterium tumefaciens; perform Agrobacterium-mediated rice genetic transformation on mature rice seeds , cultivate the regenerated plant culture; finally carry out the staining verification.

[0055] It...

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Abstract

The invention provides a rice stamen-specific expression promoter OsAnth2 and application thereof. The invention also provides an expression cassette and a recombinant expression vector containing the promoter. Specifically, the present invention applies the above-mentioned promoters to crop genetic engineering. The rice stamen-specific expression promoter OsAnth2 of the invention can regulate the concentrated expression of the gene in the stamen part of the plant, while basically not expressing in other parts. The promoter provided by the invention can improve the traits of stamens, is used for the fertility research of rice, and enriches the available genetic resources for the fertility research. In addition, crop varieties can be genetically modified through the promoter, for example, the promoter can be combined with the male sterility gene and constructed in a recombinant expression vector to breed male sterile rice varieties, and this rice variety The introduced male sterility gene is only expressed in the stamens, and will not be expressed in other parts, which is highly targeted, and can reduce the cost of biological metabolism, making it easier to ensure the stability of the transgenic variety.

Description

technical field [0001] The invention relates to the technical fields of biotechnology and crop genetic engineering. Specifically, the present invention relates to a rice stamen-specific expression promoter and its application. The promoter can drive the specific expression of a target gene in stamens in crop genetic engineering, so that it can be used without affecting the physiological activities of other tissue parts. The purpose of improving the fertility of rice by changing the structural characteristics of stamens is achieved. Background technique [0002] A promoter is a DNA sequence for the localization of RNA polymerase, usually located upstream of the coding region of a gene. Once RNA polymerase locates and binds to the promoter, the transcription process can be initiated, and its interaction with trans elements such as RNA polymerase and other protein cofactors is the essence of the regulation mode of gene expression. There are usually some special DNA sequences ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/82C12N15/84A01H5/00A01H6/46
CPCC07K14/415C12N15/8289
Inventor 魏鹏程杨剑波李莉李浩李娟杨亚春许蓉芳秦瑞英
Owner RICE RES ISTITUTE ANHUI ACAD OF AGRI SCI