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Regulatory factor for anti-reverse transcription of corn, and its coding gene and application thereof

A regulatory factor and reverse transcription technology, applied in the fields of application, genetic engineering, plant gene improvement, etc., to achieve the effect of increasing yield

Inactive Publication Date: 2006-09-13
THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the trans factors that interact with LTRE cis elements have not been reported in monocots, especially in the important crop maize.

Method used

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  • Regulatory factor for anti-reverse transcription of corn, and its coding gene and application thereof
  • Regulatory factor for anti-reverse transcription of corn, and its coding gene and application thereof
  • Regulatory factor for anti-reverse transcription of corn, and its coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1, the acquisition of LBF gene

[0035] 1. Construction of maize immature embryo cDNA library

[0036] 1. Extraction of total RNA and isolation of mRNA from immature maize embryos

[0037] Get 1g of immature embryos on the 17th day (17dpp) after pollination of maize variety Qi 319, use RNAgents Total RNAIsolation System kit (Promega Company) to extract the total RNA of the above-mentioned immature embryos of maize, then use PolyATtract mRNA Isolation System (Promega Company) from the above-mentioned mRNA was isolated from total RNA from immature maize embryos.

[0038] 2. Construction of maize 17dpp immature embryo cDNA library

[0039] Using the mRNA of maize 17dpp immature embryos obtained in step 1 as a template, use GibcoBRL's SuperScriptTMPlasmid System for cDNA Synthesis and Plasmid Cloning kit and operate strictly according to the kit instructions to construct a cDNA library of maize 17dpp immature embryos, and finally the library capacity is 5.2×10...

Embodiment 2

[0050] Embodiment 2, the binding specificity analysis of LBF and LTRE

[0051] 1. Binding specificity analysis of LBF and LTRE in yeast cells

[0052] The recombinant vectors pL4 and pmL4 were transformed into yeast yWAM2 (Leu - , His - ,Trp - ), obtain the yeast strain yWAM2 / pL4 integrated with pL4 and the yeast strain yWAM2 / pmL4 integrated with pmL4, then transform the plasmid containing the LBF gene obtained through the screening library in Example 1 into the recombinant yeast yWAM2 / pL4 and yWAM2 / pmL4, then at 28°C, in His - Cultured on selection medium for 3-5 days, the results are as follows: figure 1 As shown (A in the figure is the recombinant yeast yWAM2 / pL4, and B in the figure is the recombinant yeast yWAM2 / pmL4), only the recombinant yeast yWAM2 / pL4 can grow, but the recombinant yeast yWAM2 / pmL4 cannot grow, indicating that LBF can specifically Combined with the LTRE element, thereby activating the expression of the reporter gene HIS3 gene, so it can be expres...

Embodiment 3

[0062] Example 3, DNA core sequence analysis of LBF combined with LTRE

[0063] Site-directed mutations were performed on the possible core sequence "GCCGACC" of LTRE, and the mutation sequences of m1-m7 were as follows (underlined bases are mutation sites):

[0064] LTRE: 5'-ATTTCATG GCCGACC TGCTTTTT-3'(CK)

[0065] m1: 5'-ATTTCATG a CCGACCTGCTTTTT-3'

[0066] m2: 5'-ATTTCATGG t CGACCTGCTTTTT-3'

[0067] m3: 5'-ATTTCATGGC t GACCTGCTTTTT-3'

[0068] m4: 5'-ATTTCATGGCC a ACCTGCTTTTT-3'

[0069] m5: 5'-ATTTCATGGCCG g CCTGCTTTTT-3'

[0070] m6: 5'-ATTTCATGGCCGA t CTGCTTTTT-3'

[0071] m7: 5'-ATTTCATGGCCGAC t TGCTTTTT-3 ' carries out EMSA experiment with the method identical with embodiment 2, and the result is as follows image 3 As shown, lanes 1-7 are the combination products of LBF and m1, m2, m3, m45, m5, m6, and m7 probe sequences respectively, and lane CK is the control, indicating the combination products of LBF and LTRE probe sequences, indicating that LBF ...

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Abstract

The invention discloses the corn resisting transcription and adjusting factor and coding gene and appliance. The corn resisting transcription adjusting factor is among the proteid of the following amino acid residue: 1) the SEQ ID No:1 of the sequence list; 2)the amino acid residue sequence of the SEQ ID No:1 of the sequence list are replaced, appended by one to ten amino acid residue and the proteid acting with the LTRE arranging component to improve the capability of resisting contradictorily of the plane. The adjusting factor can be acted on the LTRE arranging component of the several resisting abiotic adversity dependency gene promoter zone and adjust several the expression of the resisting abiotic adversity dependency gene and improve the ability of the plane resisting the abiotic adversity such as the drought, the low temperature and the salting. So the LBF is important to breed the resisting athwart plant species particularly the corn and improve the yield of the crop particularly the corn.

Description

technical field [0001] The present invention relates to stress-related transcriptional regulatory factors in plants and their coding genes and their applications, in particular to a transcriptional regulatory factor and its coding genes for resistance to abiotic stresses such as low temperature, drought, and salinity in maize, and their use in cultivating stress resistance Improving applications in plants. Background technique [0002] Chilling injury is a natural disaster that often occurs in agricultural production, which greatly limits the planting of early spring crops and the harvest of late autumn crops. [0003] Studies have shown that under low temperature conditions, plants will actively mobilize their internal defense systems to resist external adversity stress. At this time, great changes will occur in plants, such as the synthesis of heart protein, the transformation of metabolism, and the accumulation of anti-adversity substances et al. (Hans J, Adaptations to ...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H1/00
Inventor 王磊范云六赵军
Owner THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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