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Protein IbEGF and protein related biomaterials and applications of protein and protein related biomaterials

A biomaterial and protein technology, applied to protein IbEGF and related biomaterials and application fields, can solve the problems of plant drought resistance genetic engineering application limitations, lack of plant drought resistance genes, and inability to improve plant yield and quality molecular design and other problems

Active Publication Date: 2020-07-07
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far, the drought resistance genes obtained in plants are still too few to meet the needs of molecular design for improving plant yield and quality
Therefore, the application of plant drought-resistant genetic engineering in agricultural production and commerce is greatly limited.

Method used

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  • Protein IbEGF and protein related biomaterials and applications of protein and protein related biomaterials
  • Protein IbEGF and protein related biomaterials and applications of protein and protein related biomaterials
  • Protein IbEGF and protein related biomaterials and applications of protein and protein related biomaterials

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0125] Embodiment 1, the acquisition of IbEGF gene

[0126] 1. Acquisition of cDNA template

[0127] The total RNA of the sweet potato line Xushu 55-2 tube seedlings was extracted with a plant total RNA extraction kit, and the first-strand cDNA was reverse-transcribed from the total RNA with a PrimeScriptTM RT reagent Kit with gDNA Eraser kit.

[0128] 2. Using the cDNA obtained in step 1 as a template, further design and artificially synthesize primers O-F and O-R according to the spliced ​​candidate IbEGF gene sequence, and use the cDNA obtained in step 1 as a template to perform PCR amplification to obtain a PCR amplification product of about 252bp and sequenced.

[0129] O-F: 5′-ATGGCCTCCCATAATGCTT-3′

[0130] O-R: 5′-TCATGTCGATTCAACCGACT-3′

[0131] The results show that the nucleotide sequence of the PCR amplification product is as shown in the 1st to 252nd positions from the 5' end of SEQ ID NO.2, the gene is named IbEGF gene, and the protein encoded by it is named I...

Embodiment 2

[0132] Example 2. Application of IbEGF protein in improving plant drought resistance.

[0133] 1. Construction of recombinant plasmid pCAMBIA super1300-IbEGF-GFP

[0134] 1. Artificially synthesize the double-stranded DNA molecule shown in positions 1 to 249 from the 5' end of SEQ ID NO.1. Using the double-stranded DNA molecule as a template, OE-F-Xba I: 5'-GC TCTAGA ATGGCCTCCCATAATGCTT-3' underlined is the recognition sequence of restriction endonuclease Xba I) and OE-R-Pst I: 5'-AA CTGCAG TGTCGATT CAACCGACT-3' (the underline is the recognition sequence of the restriction endonuclease Pst I) is used as a primer for PCR amplification to obtain a double strand containing the restriction endonuclease XbaI at the N-terminus and the restriction endonuclease PstI at the C-terminus DNA molecule.

[0135] 2. Digest the vector pCAMBIA super1300-GFP with restriction endonucleases Xba I and Pst I, and recover the vector backbone 1 of about 10783 bp.

[0136] 3. The double-strande...

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Abstract

The present invention discloses protein IbEGF and protein related biomaterials and applications of the protein and protein related biomaterials. The present invention firstly discloses the protein, the protein is the protein with an amino acid sequence shown in SEQ ID NO.1 or a fusion protein obtained by conjugating a protein tag with a N-terminus or / and a C-terminus of the amino acid sequence shown in the SEQ ID NO.1. The present invention further discloses the protein related biomaterials and the applications of the protein and protein related biomaterials. The present invention discloses the IbEGF protein and a coding gene thereof, the coding gene of the IbEGF protein is introduced into sweet potatoes to obtain a transgenic sweet potato plant that overexpresses the IbEGF, the transgenicplant is subjected to a drought stress treatment and strong in drought resistance, and the protein IbEGF has important application value in research of plant drought resistance.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to protein IbEGF and related biological materials and applications thereof. Background technique [0002] Abiotic stress has seriously harmed China's agriculture. Drought and soil salinization are the main factors limiting the sustainable development of China's agriculture, and they are two important environmental factors that lead to crop yield reduction or even extinction. The arid area in China accounts for 50% of the country's land area. Therefore, cultivating drought-resistant varieties is an effective way to resist drought and ensure yield and quality. Transgenic molecular breeding technology is an important means to improve plant resistance to stress through molecular means under stress conditions. This method has the advantages of fast breeding speed, easy to obtain positive plants, and good genetic stability, and has become the main means to obtain high-resistant pl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/82A01H6/00
CPCC07K14/415C12N15/8243C12N15/8251C12N15/8273
Inventor 刘庆昌翟红何绍贞赵宁周媛媛
Owner CHINA AGRI UNIV