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Rice constitutive expression promoter Os02g0752800 and application

A constitutive expression and promoter technology, applied in the field of plant genetic engineering, can solve the problems that the number of genes cannot match the growth of the number of constitutive promoters, and the lack of constitutive promoters, etc., to achieve the effect of long-lasting expression and wide application prospects

Active Publication Date: 2021-03-09
HEBEI UNIVERSITY OF SCIENCE AND TECHNOLOGY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

A large number of excellent functional genes have been obtained from rice, but there are relatively few reports on constitutive promoters, and the number is far from matching the increase in the number of genes

Method used

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  • Rice constitutive expression promoter Os02g0752800 and application
  • Rice constitutive expression promoter Os02g0752800 and application
  • Rice constitutive expression promoter Os02g0752800 and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Obtaining of rice constitutive promoter OsP02g0752800

[0033] 1. Design of primers

[0034] According to the whole genome sequence of the rice variety Nipponbare (Oryza sativa L cv. Nipponbare) provided by NCBI, the amplification primers were designed according to the sequence of the rice OsP02g0752800 promoter, the PCR primers were designed, the upstream primer was added with the AAGCTT restriction site of HindIII, and the downstream primer BamHI The GGATCC restriction site. The primer sequences are as follows:

[0035] Upstream primer (SEQ ID No: 2):

[0036] 5'-ATAT AAGCTT ATCACAAATGCAGGAGATGGAT-3';

[0037] Downstream primer (SEQ ID No: 3):

[0038] 5'-CG GGATCC CGGCAAGAATCCAGAAGAGAA-3'.

[0039] 2. PCR amplification

[0040] Use the rice Nipponbare genomic DNA extracted from Quanshijin Plant Genome Kit as a template, and perform PCR amplification with the upstream and downstream primers designed in step 1 to obtain PCR products.

[0041]The PC...

Embodiment 2

[0045] Example 2 Functional verification of the Os02g0752800 promoter

[0046] 2.1 Obtaining of transgenic strains

[0047] (1) Construction of expression vector

[0048] The recombinant vector pTOPOCloning Vector-OsP02g0752800 obtained in Example 1 was cut with restriction endonuclease Hind III and BamH I to obtain the OsP02g0752800 fragment; the pCAMBIA1391Z vector was obtained with restriction endonuclease Hind III and BamH I to obtain the pCAMBIA1391Z vector Skeleton fragment; the OsP02g0752800 fragment is connected with the pCAMBIA1391Z vector backbone fragment to obtain the recombinant plasmid pCAMBIA1391Z-OsP02g0752800( figure 2 ), and sequenced.

[0049] The recombinant plasmid pCAMBIA1391Z-OsP02g0752800 with correct sequencing is a vector obtained by inserting the OsP02g0752800 promoter shown in SEQ ID No: 1 between the Hind III and BamH I restriction sites of the pCAMBIA1391Z vector, and keeping other sequences of the pCAMBIA1391Z vector unchanged; OsP02g0752800 ...

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Abstract

The invention discloses a rice constitutive expression promoter Os02g0752800 and application, and relates to the technical field of plant genetic engineering. The rice constitutive expression promoterOs02g0752800 is derived from a rice japonica rice variety Nipponbare, and the size of a promoter fragment is 2170 bp. The rice constitutive expression promoter Os02g0752800 has the following characteristics that (1) the rice constitutive expression promoter Os02g0752800 is located at the 5' end and the upstream of an Os02g0752800 gene; (2) the length of a basic group is 2170bp; (3) the rice constitutive expression promoter Os02g0752800 has a necessary site for initiating transcription and a transcription starting point; and (4) the rice constitutive expression promoter Os02g0752800 can be expressed in all tissues and organs of rice. The rice constitutive expression promoter Os02g0752800 disclosed by the invention can be effectively expressed in all the tissues and organs, and the functions of an exogenous gene expression product can be fully exerted, so that an exogenous gene is efficiently, stably and durably expressed in plants.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering, in particular to rice constitutive expression promoter Os02g0752800 and its application. Background technique [0002] The growth and development of higher plants is the result of the orderly expression and synergy of different genes in different time and space. Promoter is an important cis-acting element of gene expression, which determines the expression pattern and expression intensity of genes by combining with transcription factors. Promoters can be divided into three types: constitutive, inducible, and tissue-specific promoters according to their modes of regulating gene transcription. [0003] Constitutive promoters are the earliest and most widely used promoters in plant genetic engineering. In transgenic breeding or for the study of gene function, in order to give full play to the functions of the expression products of foreign genes, constitutive promoters are genera...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/82A01H5/00A01H6/46
CPCC12N15/8222C07K14/415Y02A40/146
Inventor 艾鹏飞巴斅轲张治国
Owner HEBEI UNIVERSITY OF SCIENCE AND TECHNOLOGY
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