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.
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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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