A multi-stress-inducible promoter pg5 derived from Eucalyptus grandis and its application
A technology of promoter and Eucalyptus grandis, applied in the field of genetic engineering, can solve problems such as inhibiting plant growth and development, and achieve the effect of saving money and improving the ability to resist various adversities
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0032] promoter discovery
[0033] Genomic DNA of Eucalyptus grandis (variety EG6) was extracted, and the extraction of Eucalyptus genomic DNA referred to conventional methods in the art. Design specific primers (P1: 5'-TCAT TCTAGA GCGTAATTGCTGTCGCACCC-3', such as SEQ ID NO: 2, the underline is the XbaI restriction site) and (P2: 5'-GCTC AAGCTT CGAGAAGGAATCGCTTCTGA-3', such as SEQ ID NO: 3, the underline is the SacI restriction site) to amplify the genomic DNA, recover and clone the fragment, and perform DNA sequencing to obtain a 720bp DNA fragment, the nucleotide of the 720bp DNA fragment The sequence is shown in SEQ ID NO:1. The amplified sequence shown in SEQ ID NO: 1 was analyzed for the regulatory region, the start codon ATG of the gene was identified, and the part of the structural gene was removed. Obtain the regulatory sequence at the 5' end of the gene, the regulatory elements of the promoter such as figure 2As shown, the TATA box is located in the region?150-...
Embodiment 2
[0035] 1. Low temperature and drought-induced activity verification of the promoter pG5
[0036] Construction of recombinant expression vector
[0037] 1. Digest pGEM-T: pG5 prepared in Example 1 with restriction endonucleases XbaI and SacI, and recover the digested product (pG5).
[0038] 2. Digest the binary vector pBI121 with restriction endonucleases XbaI and SacI to obtain the pBI121 vector backbone with the CaMV35S promoter removed.
[0039] 3. Ligate the enzyme-digested product of step 1 with the vector backbone of step 2 to obtain a recombinant plasmid, and perform sequencing verification. The sequencing results show that the target plasmid pG5-GUS (replacing the XbaI of pBI121 with pG5 of the sequence shown in SEQ ID NO: 1) and the small fragment between the SacI recognition site).
[0040] 2. Transformation of tobacco by transient expression
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More 