Safflower MYB (v-myb myeloblastosis viral oncogene homolog (avian)) gene and application thereof

A gene, safflower technology, applied in the application, genetic engineering, plant genetic improvement and other directions

Inactive Publication Date: 2015-08-19
JILIN AGRICULTURAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, studies on safflower MYB, which regula

Method used

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  • Safflower MYB (v-myb myeloblastosis viral oncogene homolog (avian)) gene and application thereof
  • Safflower MYB (v-myb myeloblastosis viral oncogene homolog (avian)) gene and application thereof
  • Safflower MYB (v-myb myeloblastosis viral oncogene homolog (avian)) gene and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1 safflower ( Carthamus tinctorius L .) Petal RNA extraction and cDNA synthesis

[0025] RNA was extracted from Tacheng safflower petals with TRIZOL Plus, and 1 μg of total RNA was reverse-transcribed into a cDNA template.

Embodiment 2

[0026] Verification of the sequence fragment of embodiment 2 MYB gene

[0027] According to the candidate genes selected from the sequencing results of safflower 454 in our laboratory, specific primers (Table 1) were designed to verify the gene fragments. According to the verified gene fragments of MYB, primers (Table 1) were designed for gene fragment cloning. According to splicing and alignment of safflower 454 sequencing results, the sequence fragment of MYB gene was obtained, and a 798bp fragment containing the start codon was amplified by RT-PCR ( figure 1 A), RT-PCR reaction system is shown in Table 2. The recovered fragments were connected to pEASY-T1 (Beijing Quanshijin Biotechnology Co., Ltd.), and further passed bacterial liquid PCR ( figure 1 B) and ECO RI and Hin dⅢ enzyme digestion identification ( figure 1 C), the sequencing result is correct.

[0028]

Embodiment 3

[0029] Example 3 Cloning of full-length cDNA of MYB gene

[0030] According to the 798bp fragment sequence of the MYB gene, a pair of primers (Table 1) were designed, and the cDNA of the safflower petal was used as a template for 3' RACE cloning to obtain the 3' end sequence of the safflower MYB gene. The full-length sequence of safflower MYB12 was obtained by splicing, the full-length primers (Table 1) were redesigned, and PCR amplification was performed to obtain the full-length cDNA of the safflower MYB12 gene ( figure 2 ),named Ct-MYB12 . Compared with the sequence on NCBI, it was found that the homology of this gene and dahlia MYB12 reached 80%, indicating that the gene is indeed the MYB gene of safflower.

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Abstract

The invention discloses a safflower MYB (v-myb myeloblastosis viral oncogene homolog (avian)) gene. The safflower MYB gene has the advantages that the expression characteristic of a transcriptional modulatory gene is realized; by utilizing the transcription to modulate the expression of the related flavone metabolism gene, the content of total flavone in the safflower is improved, and a firm foundation is laid for culturing new products with high flavone contents.

Description

technical field [0001] The invention belongs to the field of bioengineering, and in particular relates to the MYB12 gene of safflower and its application in cultivating transgenic plants with high flavonoid content. Background technique [0002] safflower( Carthamus tinctorius L.) is an annual herbaceous plant belonging to Compositae and Safflower. Safflor yellow (safflor yellow), as its main active ingredient, is a mixture of water-soluble chalcones isolated from its dried tubular flowers. Blood pressure, cerebral infarction, anti-tumor, diabetic complications, chronic kidney disease and other diseases. It can be seen that safflower yellow is a natural medicine with great potential. Although the planting area of ​​safflower continues to expand in my country, it still fails to solve the problem of scarcity of safflower yellow pigment resources. It is of great economic significance to increase the yellow pigment content of safflower itself and cultivate new lines of saff...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/82A01H5/00
Inventor 官丽莉李海燕李校堃杨晶杜林娜王法微董园园
Owner JILIN AGRICULTURAL UNIV
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