Soybean folic acid synthesis key enzyme ADCS and gene and application thereof

A key enzyme gene, key enzyme technology, applied in soybean folic acid synthesis key enzyme ADCS and its gene and application field

Active Publication Date: 2013-01-09
THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

According to the report of the United States Department of Agriculture (2001), compared with cereals, fruits, vegetables, and dairy products, soybean folic acid content is the highest, which indicates that there may be ADCS genes in soybeans, but there is still no information about the cloning of ADCS genes in soybeans so far. not reported

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Soybean folic acid synthesis key enzyme ADCS and gene and application thereof
  • Soybean folic acid synthesis key enzyme ADCS and gene and application thereof
  • Soybean folic acid synthesis key enzyme ADCS and gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0166] Cloning and sequence analysis of embodiment 1, GmADCS gene

[0167] 1. Obtain the EST sequence of soybean ADCS gene

[0168] The CDS sequence (AtADCS) of the Arabidopsis ADCS gene (AtADCS), with a size of 2760bp, was compared with the EST sequence of soybean through NCBI. There were 8 soybean EST sequences, which were highly consistent with the 2050-2714bp of AtADCS, and were named 3 respectively. 'EST1-8, an EST sequence with a size of 281bp, which is highly consistent with AtADCS's 1345-1592bp, named mEST. And the 8 sequences of 3'EST1-8 were compared and found that there were only differences in individual bases and sequence lengths between them. After analysis, the downstream primer 3 was designed using the 3'EST5 terminal sequence with high homology as a template 'EST5-R1 / R2, using the mEST sequence as a template to design the upstream primer mESTF1 (see Table 1 for the primer sequence).

[0169] The sequences of 3'EST1, 3'EST2, 3'EST3, 3'EST4, 3'EST5, 3'EST6, 3'...

Embodiment 2

[0181] Embodiment 2, the expression pattern of GmADCS gene in soybean

[0182] The transcription level of GmADCS gene in different tissues and organs of soybean was analyzed by RT-PCR. Soybean roots, stems, and leaves grown for 8 weeks were taken, and total RNA was extracted by the Trizol method, reverse-transcribed into cDNA as a template, and GmActin-specific primers GmActin11F, GmActin11R (primer See Table 2 for the sequence), design gene-specific primers GmADCS-FSP and GmADCS-RSP (see Table 2 for the primer sequences) according to the CDS sequence of the GmADCS gene, and the results show that the gene can be transcribed in roots, stems and leaves, wherein, Expression levels are highest in leaves and weaker in roots and stems (e.g. figure 2 shown).

[0183] Table 2. Gene-specific primers for GmACTIN and GmADCS

[0184] Primer name

Embodiment 3

[0185] Example 3, the acquisition of transgenic GmADCS gene Arabidopsis and the determination of its folic acid synthesis precursor pABA content

[0186] The CDS sequence of GmADCS was constructed between the restriction sites NcoI and Pml I of the plant overexpression vector pCAMBIA1302, and the recombinant expression vector pCAMBIA1302-GmADCS was obtained driven by the cauliflower mosaic virus (CaMV) 35S promoter.

[0187] The above-mentioned recombinant vector was transformed into Agrobacterium LBA4404 by electroporation, and the correct clones identified by enzyme digestion were picked, and the wild-type Arabidopsis was infected by the Agrobacterium-mediated flower dipping method, and the harvested seeds were recorded as the T0 generation. The seeds of the T0 generation were first screened on the resistance medium of 1 / 2MS+hygromycin B (25mg / L), and 13 positive green seedlings (referred to as the T1 generation) were obtained, and moved into the greenhouse for 20 days, and t...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention relates to the plant genetic engineering field, in particular to a soybean folic acid synthesis key enzyme ADCS and a gene and application thereof. The amino acid sequence of the soybean folic acid synthesis key enzyme ADCS is shown in SEQ ID No.1 through the gene expression of the soybean folic acid synthesis key enzyme ADCS, the content of the folic acid synthetic precursor p-aminobenzoic acid (pABA) in the transgenic plant can be significantly increased. Therefore, the soybean folic acid synthesis key enzyme ADCS lays a foundation for the increase of the folic acid content of the plant, ensures that human can directly obtain natural folic acid through diet to meet daily need and has a significance to relieve the folic acid lack of human.

Description

technical field [0001] The invention relates to the field of plant genetic engineering, in particular to the soybean folic acid synthesis key enzyme ADCS and its gene and application. Background technique [0002] Folic acid (folates) is the general term for tetrahydrofolate (THF) and its derivatives. It is a class of water-soluble B vitamins, also known as vitamin B9, vitamin B11, and vitamin M. Its chemical structure is as follows: [0003] [0004] Among them, THF consists of three parts: a pteridine ring (2-amino-4-hydroxy-6-methylpteridine), pABA p-aminobenzoic acid, and poly-glutamic acid (poly-Glu) (1-8 A Gluλ link), also known as pteroylglutamic acid. One-carbon units of different oxidation states, such as methyl, methylene, methylene, formyl, attached to N5 of the pteridine ring, N10 of pABA or bridged both to form tetrahydrofolate (THF4), 5 -Methyl THF4, 5-formyl THF4, 5,10-methylene THF4 various folic acid derivatives. THF and its derivatives, as the donor a...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/00C12N15/52C12N15/63A01H5/00
Inventor 张春义梁秋菊范云六
Owner THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap