cDNA sequence of MYB transcription factors for positively regulating anthocyanin synthesis

A technology of transcription factors and anthocyanins, applied in genetic engineering, plant genetic improvement, plant peptides, etc.

Inactive Publication Date: 2015-12-09
NORTHEAST NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] So far, people have cloned R2R3MYB that positively regulates anthocyanin synthesis in many plants, such as snapdragon, Arabidopsis, cabbage, pepper, sweet orange, epimedium, mangosteen, gentian, gerbera, sweet potato, Cow, apple, alfalfa, bayberry, tobacco, petunia, pear, sand pear, peach, tomato, potato and grape, etc., but these are all dicotyledonous plants, and only clones from lily, phalaenopsis and corn in monocotyledons The MYB transcription factor related to anthocyanin synthesis was obtained

Method used

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  • cDNA sequence of MYB transcription factors for positively regulating anthocyanin synthesis
  • cDNA sequence of MYB transcription factors for positively regulating anthocyanin synthesis
  • cDNA sequence of MYB transcription factors for positively regulating anthocyanin synthesis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] [Example 1-1] Obtaining the target fragment: according to the Arabidopsis PAP1 sequence, perform local Blast. The sequence with high homology to Arabidopsis PAP1 was screened, and bioinformatics analysis was performed to obtain a fragment with a length of 212bp, which contained the start codon ATG.

[0030] [Example 1-2] FhPAP1L1 Obtaining the 3' sequence of the gene: according to the transcriptome sequencing sequence, design a specific upstream primer, namely 5'-GGCAGGACTAAGAAGGTGTCG-3'; design a specific downstream primer according to the conserved polyA structure of mRNA: 5'-AAAAAAAAAAAAAAAATCGATGTCGACTCGAGTC-3'. Trizol kit was used to extract the total RNA in the blooming period of the safflower safflower, reverse transcribed to generate the first strand of cDNA, and used this as a template for RACE to obtain a 717bp PCR product.

Embodiment 1-2

[0031] [Example 1-3] FhPAP1L1 Obtaining the full-length sequence of the gene: According to the results obtained from the above sequencing, design specific upstream primers, namely, 5'-CCTCCCCCTAATGTCTTTATCTTG-3' and 5'-ATATATAGATATCAGTATTGTTA-3' downstream primers. Use the Trizol kit to extract the total RNA of the petals of Cymbidium safflower, and reverse transcribe to generate the first strand of cDNA, which is used as a template for RT-PCR to obtain FhPAP1L1 The full-length sequence of the gene.

Embodiment 1-3

[0032] Example 2 FhPAP1L2 Cloning of full-length genes

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Abstract

The invention discloses a cDNA sequence of two MYB transcription factors (FhPAP1L1 and FhPAP1L2) which are cloned from petals of freesia refracta for the first time and are used for positively regulating anthocyanin synthesis, and a coded amino sequence is obtained through conjecture. Through over-expression of FhPAP1L1 and FhPAP1L2 in arabidopsis thaliana and tobacco, coloring of plant tissues can be significantly enhanced, so that the function of the FhPAP1L1 and FhPAP1L2 of participating in the positive regulation for plant anthocyanin synthesis is determined.

Description

technical field [0001] The invention belongs to the technical field of plant molecular biology, in particular to two functional MYB transcription factors ( FhPAP1L1 and FhPAP1L2 ) cDNA sequence and its amino acid sequence. Background technique [0002] There are three main groups of pigments that affect the color of plants: anthocyanins, carotenoids and betalains. People have great interest in the study of anthocyanins, which stems from their important roles in plant physiology and human health. Anthocyanins are catalyzed and synthesized by a series of enzymes, which are controlled by corresponding structural genes. Studies have shown that the expression of these structural genes is regulated by a complex transcriptional regulatory network, in which the MYB transcription factor is MBW (MYB-bHLH-WDR) An important member of the transcription factor complex, which plays a role in specific regulation. The plant MYB transcription factor family is a large family with differen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415
Inventor 高翔王丽李月庆单晓彤
Owner NORTHEAST NORMAL UNIVERSITY
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