Protein IbEGF and related biological material and application thereof

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, inability to meet the molecular design for improving plant yield and quality, and lack of plant drought resistance genes.

Pending Publication Date: 2020-06-12
CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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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 yiel...

Method used

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  • Protein IbEGF and related biological material and application thereof
  • Protein IbEGF and related biological material and application thereof
  • Protein IbEGF and related biological material and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0125] Example 1. Obtaining IbEGF gene

[0126] 1. Obtaining cDNA template

[0127] The total RNA of the test tube plantlets of the sweet potato line Xushu 55-2 was extracted with the plant total RNA extraction kit, and the first strand cDNA was reverse transcribed from the total RNA using 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 OF and OR according to the candidate IbEGF gene sequence of the splicing. Using the cDNA obtained in step 1 as a template, 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 showed that the nucleotide sequence of the PCR amplified product is as shown in SEQ ID NO. 2 from positions 1 to 252 from the 5'end. The gene was named IbEGF gene, and the protein encoded by it was named IbEGF protein or protein...

Embodiment 2

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

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

[0134] 1. Artificially synthesize the double-stranded DNA molecule shown in SEQ ID NO.1 from position 1 to position 249 from the 5'end. 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 TGTCGATTCAACCGACT-3' (underlined is the recognition sequence of restriction enzyme Pst I) is the primer for PCR amplification, and the double-stranded DNA containing restriction enzyme XbaI at the N-terminal and restriction enzyme Pst I at the C-terminal is obtained molecular.

[0135] 2. The vector pCAMBIA super1300-GFP was digested with restriction enzymes Xba I and Pst I, and the vector backbone 1 of about 10783 bp was recovered.

[0136] 3. Cut the double-stranded DNA molecule...

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Abstract

The invention discloses a protein IbEGF and a related biological material and application thereof. The invention firstly discloses a protein which is a protein with an amino acid sequence shown as SEQID NO.1 or a fusion protein obtained by connecting an N terminal or/and a C terminal of the amino acid sequence shown as SEQ ID NO.1 with a protein tag. The invention further discloses a biological material related to the protein and application. The IbEGF protein and the coding gene thereof are discovered, the coding gene of the IbEGF protein is introduced into sweet potatoes to obtain transgenic sweet potato plants over-expressing IbEGF, the transgenic plants are subjected to drought stress treatment, and then the drought resistance of the transgenic lines is enhanced. Therefore, the IbEGFprotein and the coding gene thereof have important application value in the research of improving drought resistance of plants.

Description

Technical field [0001] The present invention belongs to the field of biotechnology, and specifically relates to protein IbEGF and related biological materials and applications. Background technique [0002] Abiotic stress has caused serious damage to China's agriculture. Among them, drought and soil salinization are the main factors that limit the sustainable development of China's agriculture, and they are two important environmental factors that cause crop production to decrease or even cease. China's drought area 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 adversity through molecular means under stress conditions. This method has the advantages of fast breeding, easy to obtain positive plants, and good genetic stability. It has become the main means to obtain high-r...

Claims

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Application Information

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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
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