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bzip transcription factor related to corn kernel growth, its coding gene and gene expression

A technology for transcription factors and coding genes, which can be used in genetic engineering, plant genetic improvement, fermentation, etc.

Inactive Publication Date: 2008-01-02
SHANGHAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, in maize, especially maize kernels, there are still a large number of bZIP transcription factors that have not been discovered and studied in depth

Method used

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  • bzip transcription factor related to corn kernel growth, its coding gene and gene expression
  • bzip transcription factor related to corn kernel growth, its coding gene and gene expression
  • bzip transcription factor related to corn kernel growth, its coding gene and gene expression

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Obtaining a new gene of bZIP transcription factor expressed during corn kernel development

[0026] 1. Construction of a homogenized cDNA library at corn kernel development stage

[0027] In order to obtain the expression sequence of the entire grain development stage, collect corn grains 3 to 36 days after pollination, mix them in equal volumes, and extract total RNA (Trizol kit, Qiagen) to obtain the PolyATtract from Promega R The mRNA purified by the mRNA Isolation System III Kit is used as a template, and a cDNA homogenization scheme based on the principle of renaturation kinetics is used, that is, through denaturation and renaturation operations, the abundance of cDNA tends to be consistent, and finally a homogenized cDNA library is established .

[0028] 2. Acquisition of genes encoding bZIP transcription factors

[0029]The sequencing universal primer T3 / T7, dideoxy method (DYEnamic ET-Terminator Cycle Sequencing), and DNA sequence analyzer (MegaBACE 4500)...

Embodiment 2

[0030] Example 2: Identification of bZIP transcription factors

[0031] Use MEME analysis software to analyze the conserved segments of these 13 bZIP transcription factors, see Figure 2. They all contain a conservative alkaline leucine region, and MSBZIP7-12 except for the basic segment specific to bZIP protein , There is also a conservative segment with unknown function. According to the number and types of conserved segments, we divide these 13 transcription factors into 3 categories, among which MSBZIP1-6 belongs to the first category; MSBZIP7-11 belongs to the second category; and the third category includes MSBZIP12 and MSBZIP13.

[0032] Use Megabase3.0 software to cluster these 13 bZIP transcription factors, see Figure 3. The results show that MSBZIP5-11 are relatively close in evolutionary distance, while MSBZIP1-4, MSBZIP12, MSBZIP13 and O2 are relatively close in distance. It may reveal two evolutionary patterns in these bZIP transcription factors.

Embodiment 3

[0033] Example 3: Expression profile analysis of bZIP transcription factor

[0034] Using maize kernel RNA from 3-36 days after pollination as template, Oligo(dT) 16 Used as a primer for reverse transcription to obtain cDNA at various time points in the entire grain development stage. Primers were designed according to the sequences of these 13 transcription factors. See Table 1. Semi-quantitative PCR was performed using the cDNA at each time point as a template. The PCR products were analyzed by 1% agarose gel electrophoresis. See Figure 4, which proves that MSBZIP2, MSBZIP4, MSBZIP6, MSBZIP7, MSBZIP8, MSBZIP10, MSBZIP11, MSBZIP12, and MSBZIP13 are continuously expressed during grain development, see Figure 4-A; while MSBZIP1, MSBZIP3, MSBZIP5, MSBZIP9 are specifically expressed during development, see Figure 4 -B.

[0035] Table 1. Primer list of bZIP transcription factor

[0036] Gene name

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Abstract

The invention relates to thirteen new genes of bzip transcription factor expressed in the shelled corn growth process and their sequence analysis and expression analysis. The name of bzip transcription factor gene is MSBZIP1-MSBZIP13, and the nucleic acid sequence is one of the below: (1)1-13 DNA sequence in the sequence table, (2)1-13 limited DNA sequence with more than 90% consanguinity in the sequence table and DNA sequence with the same coding functional protein. The protein which is coded by bzip transcription factor gene has 14-26 amino acid residue sequence in the sequence table or has the same active 14-16 derived protein sequence with 14-26 amino acid residue sequence by replacing, missing or adding 14-26 amino acid residue sequence. Thirteen new genes of bzip transcription factor in the core is reported firstly, which provides the therapy for analyzing the expression and control of the gene in the shelled corn growth process and utilizing these transcription factors.

Description

Technical field [0001] The present invention relates to 13 new bZIP transcription factors and their coding genes related to corn grain development, and provides the expression characteristics of these transcription factor genes during grain development. technical background [0002] Corn (Zea mays L.), also known as corn, is the third largest food crop in the world, and its sown area is second only to rice and wheat. Since 2000, the total output and yield of corn have ranked first. Corn is not only a high-yield food crop, but also a nutrient-rich feed crop and a good industrial raw material crop. Maize kernel is the most important production trait of maize. In addition to providing people with food sources, it is actually an important reproductive organ and an important model for the study of gramineous seed development. The development of corn kernels starts from fertilization. Through the process of double fertilization, one sperm released from the tip of the pollen tube fuses ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415
Inventor 宋任涛朱佳冯玲娜王刚许政暟
Owner SHANGHAI UNIV
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