Banana fruit differential high efficient starter and plant expressing carrier

A high-efficiency expression, promoter technology, applied in the direction of plant gene improvement, botanical equipment and methods, introduction of foreign genetic material using vectors, etc.

Inactive Publication Date: 2006-10-11
INST OF TROPICAL BIOSCI & BIOTECH CHINESE ACADEMY OF TROPICAL AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

According to previous studies, the promoter of the BanLec gene is likely to be a fruit-specific high-efficiency expression promoter

Method used

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  • Banana fruit differential high efficient starter and plant expressing carrier
  • Banana fruit differential high efficient starter and plant expressing carrier
  • Banana fruit differential high efficient starter and plant expressing carrier

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Embodiment Construction

[0059] The present invention will be described in detail below in conjunction with the example application (using the promoter to transform the stilbene synthase gene).

[0060] 1. Cloning of the promoter sequence: design 5' and 3' primers according to the sequencing results of the BanLec gene promoter, and add HindIII and BamHI respectively

[0061] Restriction sites:

[0062] 5'Primer: 5'-gc AAGCTT GGTATCTATTTATGAAATC--3,

[0063] 3'Primer: 3’--- gGGATC C ATCGACGTCAACACATAACG-3'

[0064] PCR amplification of the target fragment: Add in sequence to the PCR tube

[0065] Sterilized water 18.9ul

[0066] Banana genomic DNA 1.0ul

[0067] Buffer 2.5ul

[0068] dNTP 0.0ul

[0069] TaqDNA polymerase 0.5ul

[0070] 5'Premer 1.5ul

[0071] 3'Premer 1.5ul

[0072] Total volume 25ul.

[0073] Gently mix, centrifuge briefly, put the Eppendorf tube in a PCR instrument, and cover with a heating cap. Reaction procedure:

[0074] 7 cycles of 94°C 2S, 72°C 3min;

[007...

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Abstract

The invention relates to a banana fruit-specific high-efficiency expression promoter and a plant expression vector thereof. The promoter is 748 bp long, and a new plant expression vector named pBIL2 is constructed by replacing the constitutive 35S promoter on the plant expression vector pBI121. The GUS gene transient expression method was used to detect the promoter activity of the promoter containing this segment and the pBI121 promoter. GUS tissue staining showed that the promoter had expression tissue specificity, that is, it was only expressed in fruit. The activity of GUS was determined to be 100-300 times higher than that of other organs in each banana fruit, and it was slightly higher than that of 35S promoter. The acquisition of the promoter provides a powerful tool for banana transgene and molecular biology research on banana fruit, especially creates conditions for the research and development of using banana as a bioreactor. Therefore, the acquisition of the promoter has important theoretical and practical significance.

Description

[technical field] [0001] The present invention relates to the acquisition of the banana Banlec gene promoter sequence and the construction of the plant expression vector. [Background technique] [0002] Banana (Musa spp.) belongs to Musa family (Musaceae). All edible bananas are cultivars of the genus Musa, and are important tropical and subtropical fruits. Banana meat is sweet and glutinous, fragrant and delicious, and has high nutritional value. In addition to eating fresh and cooking, it can also be processed into a variety of nutritious products. Banana is an ideal plant for producing recombinant medicinal proteins and peptides or valuable compounds by transgenic methods. Its advantages as a bioreactor mainly include the following aspects: 1. Its growth period is short, ten to twelve 2, banana is the fruit with the largest consumption of fresh fruit, fresh food can guarantee that the medicinal protein expressed by the gene in the fruit will not be denatured, and is an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N15/63
Inventor 徐碧玉金志强刘歌刘菊华
Owner INST OF TROPICAL BIOSCI & BIOTECH CHINESE ACADEMY OF TROPICAL AGRI SCI
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