Tissue-specific promoter and application thereof

A tissue-specific, promoter technology, applied in the field of indica rice ABCG15 gene promoter, can solve problems such as unresearched

Active Publication Date: 2014-04-16
BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] ABCG15 belongs to the ABC (ATP Binding cassette) transfer protein G subfamily member (Paul J.Verrier et al.Trends in Plant Science2008.13 (14): 151-159), it has been reported that the promoter of the ABCG15 gene acts as an anther tapetum tissue The specific expression promoter was isolated and identified from japonica rice, but there is no related research on the ABCG15 gene promoter derived from indica rice

Method used

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  • Tissue-specific promoter and application thereof
  • Tissue-specific promoter and application thereof
  • Tissue-specific promoter and application thereof

Examples

Experimental program
Comparison scheme
Effect test

no. 1 example

[0067] First Example, Obtaining and Morphological Observation of Osats1 Mutant Plants

[0068] The mutant plant Osats1 (obtained by the cooperation project of Sichuan Agricultural University) is a natural mutation material identified in the process of indica hybrid breeding, and the material has no significant difference from the wild type in the vegetative growth stage of the plant ( image 3 ), but the anthers are inhibited in the early stage of development, and in the later stages of flower development, the anthers appear white and thin, and eventually fail to produce pollen, showing complete male sterility ( Figure 4 ).

no. 2 example

[0069] The second embodiment, the acquisition and analysis of the anther-specific expression promoter prOsATS1

[0070] Through fine mapping, it was found that the Osats1 mutant plant was a loss-of-function mutant caused by the deletion of 12bp in the ABCG15 gene (Loc_Os06g40550; as shown in SEQ ID NO: 2 in the sequence listing). The sequence of the mutant plant Osats1 is shown in the sequence listing as SEQ ID NO: 3.

[0071] The analysis found that there is an RNA polymerase recognition site at 2066 bp upstream of the finely positioned ABCG15 gene, as shown in SEQ ID NO: 1 in the sequence listing, named prOsATS1. This promoter is specifically expressed in anthers, especially tapetum tissue.

[0072] Through NSITE-PL online prediction, it is found that prOsATS1 (SEQ ID NO: 1) includes 5 core regulatory elements (http: / / linux1.softberry.com / berry.phtml?topic=nsitep&group=programs&subgroup=promoter), the specific information is shown in the table 1:

[0073] Table 1: Predict...

no. 3 example

[0076] The third embodiment, construction of recombinant expression vector and transformation of recombinant expression vector into Agrobacterium

[0077] 1. Construction of a recombinant cloning vector containing the prOsATS1 promoter sequence

[0078] The prOsATS1 promoter sequence was connected to the cloning vector pGEM-T (Promega, Madison, USA, CAT: A3600), and the operation steps were carried out according to the instructions of the pGEM-T vector product of Promega Company to obtain the recombinant cloning vector pT-prOsATS1. The construction process is as follows: figure 1 Shown (wherein, Amp represents the ampicillin resistance gene; f1 represents the replication origin of phage f1; LacZ is the LacZ start codon; SP6 is the promoter of SP6 RNA polymerase; T7 is the promoter of T7 RNA polymerase; prOsATS1 is the promoter of prOsATS1 sequence (SEQ ID NO: 1); MCS is the multiple cloning site).

[0079] Then, the recombinant cloning vector pT-prOsATS1 was transformed into ...

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Abstract

The invention relates to a tissue-specific promoter and an application thereof. A nucleotide sequence of the tissue-specific promoter comprises a sequence represented by (a) or (b) as follows: (a) a sequence comprising nucleotide sequences represented by nucleotide at positions 920-932, nucleotide at positions 947-961, nucleotide at positions 1507-1519, nucleotide at positions 1963-1971 and nucleotide at positions 2018-2028 in SEQ ID NO:1 simultaneously; (b) a nucleotide sequence hybridized with the nucleotide sequences limited by (a) under stringent conditions. The tissue-specific promoter is isolated from an ABCG15 gene of a nonglutinous rice variety 93-11 for the first time; in addition, order-adjustable heterogeneous nucleotide sequences are specifically expressed in plant anther tissue, particularly tapetum tissue, so that a foundation is laid for creation of a novel male sterility line.

Description

technical field [0001] The invention relates to a tissue-specific promoter and its use, in particular to a promoter of the indica rice ABCG15 gene specifically expressed in anther tapetum tissue and its use. Background technique [0002] Expression of a heterologous DNA sequence in a plant host is dependent on having an operably linked promoter that is functional in the plant host. The choice of promoter sequence will determine when and where the heterologous DNA sequence is expressed in the plant host. Thus, when expression in plant-preferred tissues is desired, tissue-specific promoters are used. Conversely, when expression in whole plant cells is desired, constitutive promoters are used. For example, plant resistance to insect attack can be achieved by genetically manipulating the plant genome to contain a tissue-specific promoter operably linked to a heterologous insect-resistant gene, so that the insect-resistant substance is specifically expressed in susceptible plan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/84C12N1/21C12N5/10A01H5/00
Inventor 于彩虹
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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