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Specific promoter GmFTL2 for anthers, ovules and root caps of soybeans and application of specific promoter GmFTL2

A specific promoter, promoter technology, applied in the fields of genetic engineering and molecular biology

Active Publication Date: 2017-06-30
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few reports on promoters specifically expressed in soybean anthers, ovules and root caps

Method used

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  • Specific promoter GmFTL2 for anthers, ovules and root caps of soybeans and application of specific promoter GmFTL2
  • Specific promoter GmFTL2 for anthers, ovules and root caps of soybeans and application of specific promoter GmFTL2
  • Specific promoter GmFTL2 for anthers, ovules and root caps of soybeans and application of specific promoter GmFTL2

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 Obtaining of soybean anther, ovule and root cap-specific promoter GmFTL2

[0040] Use the following 5 pairs of primers to amplify from the soybean genome by segmented PCR and digest and connect to the entry vector PEC ( figure 2 ) (see Xu W et al., BMC Plant Biology: BioVector, a flexible system for genespecific-expression in plants), sequenced to obtain a total length of 9978bp GmFTL2 promoter, such as figure 1 Shown, its nucleotide sequence is shown in SEQ ID NO:1.

[0041] The primer sequences used are as follows:

[0042] GmFTL2-1-F: 5'-GAGAGGTCGACCATGTAAAACCTTCATTGTACTTCT-3'

[0043] GmFTL2-1-R: 5'-AAGCCGCCAACGTTATAT-3';

[0044] GmFTL2-2-F: 5'-ACAATGCATCAATATATAACGTT-3'

[0045] GMFTL2-2-R: 5'-TCTAGAAATAAAAATGAAAAAAATA-3';

[0046] GmFTL2-3-F: 5'-TAAGGGAGCCAAAATCCAACAATA-3'

[0047] GmFTL2-3-R: 5'-TTTGTAAGTCCACCTGAACCTCCAC-3';

[0048] GmFTL 2-4-F: 5'-TCAGCTAGTGCCCGTTACAG-3'

[0049] GmFTL 2-4-R: 5'-CTAGTGTTTTCTCCCTTC-3';

[0050] GmFTL 2-5-F: 5'...

Embodiment 2

[0055] Example 2 Application of soybean-specific promoter GmFTL2 in regulating the expression of downstream gene GUS

[0056] After the GmFTL2 promoter was obtained by PCR cloning, it was ligated to the entry vector PEC by enzyme digestion, and the GUS gene was ligated to another entry vector GEC at the same time, and the promoter and GUS gene were combined by LR reaction using the GateWay recombination cloning method (Invitrogen). Recombined into the expression vector BDV2. For entry vector GEC and expression vector BDV2, see Xu W et al., BMC Plant Biology: BioVector, a flexible system for gene specific-expression implants. The structures of plasmids GEC and BDV2 are as follows: image 3 , Figure 4 shown. The binary expression vector pGmFTL2::GUS was obtained; the plant expression vector pGmFTL2::GUS was transformed into Escherichia coli DH5α for propagation. The transformation method of Escherichia coli competent cells includes: (1) preparing LB solid medium containing ...

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Abstract

The invention discloses a specific promoter GmFTL2 for anthers, ovules and root caps of soybeans. The nucleotide sequence of the specific promoter GmFTL2 is shown as SEQ NO:1. The invention further provides an application of the promoter in regulation of downstream gene expression. The promoter GmFTL2 is used for overexpressing a GUS gene in arabidopsis thaliana, and results show that the promoter GmFTL2 can obviously increase expression quantity of the GUS gene in anthers, ovules and root caps of plants, so that the promoter GmFTL2 has specificity in the anthers, the ovules and the root caps of the plants and is suitable for spherically expressing a target gene in the anthers, the ovules and the root caps of the plants such as various crops, forests, vegetables, flowers, grass and the like.

Description

technical field [0001] The invention belongs to the technical fields of molecular biology and genetic engineering, and in particular relates to a soybean anther, ovule and root cap specific promoter GmFTL2 and its application. Background technique [0002] The promoter is an important cis-acting element in a gene, generally located tens of bases upstream of the transcription initiation site. According to different transcription modes, promoters can be divided into three types: constitutive promoters, tissue or organ-specific promoters, and inducible promoters. The activity of tissue- or organ-specific promoters is induced and regulated by specific tissue cell structures and chemical and physical signals, that is, the expression of such promoters has specific spatiotemporal characteristics. Driven by such promoters, the expression of genes is often limited to certain specific organs or tissue parts or specific developmental periods. Tissue or organ-specific promoters can no...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/82C12N15/11A01H5/00
CPCC07K14/415C12N15/8227C12N15/8231C12N15/8233
Inventor 傅永福张晓玫陈福禄刘丽敏
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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