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Plant pistil specific promoter and its application

A technology of plants and primer pairs, applied in the fields of applications, plant peptides, plant products, etc.

Active Publication Date: 2020-12-25
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the isolation, cloning and application of regulatory elements for organ / tissue / cell-specific gene expression will inevitably become the next unavoidable technical challenge in molecular biology and biotechnology research.

Method used

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  • Plant pistil specific promoter and its application
  • Plant pistil specific promoter and its application
  • Plant pistil specific promoter and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Embodiment 1, the acquisition of CSP1 promoter

[0049] 1. Using the genomic DNA of the rice variety Zhonghua 11 as a template, PCR amplification was performed using the pCSP1-KpnI-F and pCSP1-SmaI-R primer pairs to obtain PCR amplification products. The primer sequences are as follows:

[0050] pCSP1-KpnI-F:agggGGTACCctcttacttgggggaggcctg (SEQ ID NO: 2);

[0051]pCSP1-SmaI-R: aaaaCCCGGGatctcctccgccgccgcgaa (SEQ ID NO: 3).

[0052] 2. Perform 1% agarose gel electrophoresis detection on the PCR amplification product obtained in step 1, recover and purify the PCR amplification product. and sequence it.

[0053] The agarose gel electrophoresis of the PCR amplification product is shown in figure 1 (Swimming lane M is the DNA molecular weight standard, and each band is 5000bp, 3000bp, 2000bp, 1000bp, 750bp, 500bp, 300bp, 200bp in order from large to small; Swimming lanes 1-4 are PCR amplification products, about 1.1kb).

[0054] Sequencing results showed that a DNA frag...

Embodiment 2

[0055] Example 2, Functional Verification of CSP1 Promoter

[0056] 1. Obtaining of transgenic pCSP1:GUS rice

[0057] 1. Preparation of backbone vector pCAMBIA1305NO35S

[0058] The carrier pCAMBIA1305.1 (the structure of the carrier is as shown in Fig. Figure 6 shown) to carry out double enzyme digestion, recover the large fragments and then self-ligate to obtain the ligation product, which is the backbone vector pCAMBIA1305NO35S. The backbone vector pCAMBIA1305NO35S was transformed into Escherichia coli, and positive colonies were picked for sequencing identification.

[0059] Sequencing results showed that the backbone vector pCAMBIA1305NO35S was obtained by deleting the 35S promoter between the HindIII and NcoI restriction sites in the pCAMBIA1305.1 vector and keeping other sequences of the pCAMBIA1305.1 vector unchanged.

[0060] 2. Construction of recombinant plasmid pCSP1:GUS

[0061] 1) The pCAMBIA1305NO35S vector obtained in step 1 was double-digested with restr...

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Abstract

The invention discloses a plant pistil specific promoter and application thereof. The invention provides a CSP1 promoter which is any one of DNA molecules described in the following 1)-3): 1), a DNA molecule as shown in a sequence 1 in a sequence list; 2), a DNA molecule hybridized with the DNA molecule in 1) under stringent conditions and having a promoter function; 3), a DNA molecule having morethan 90% homology of the DNA molecule in 1) and having the promoter function. Experiments prove that CSP1 can effectively start expression of an exogenous gene and has an important application valuefor transgenic plant culturing.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a plant pistil-specific promoter and application thereof, in particular to a rice pistil-specific expression promoter and application thereof. Background technique [0002] my country is a large agricultural country, and food security affects my country's economic, social and political stability. In the medium and long term, due to the increase in population, decrease in arable land, accelerated urbanization, and improvement in people's living standards, my country's food demand will show rigid growth, and the relationship between food supply and demand will be tight. Therefore, improving crop yield and quality is an important condition to ensure the sustainable development of agriculture in our country. Crop seed size and weight are important yield traits. Finding genes related to crop seed size and weight has important theoretical significance and application value for...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/82A01H5/02A01H6/46
CPCC07K14/415C12N15/8205C12N15/8233
Inventor 王东辉白书农叶思达许智宏
Owner PEKING UNIV
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