2results about How to "High expression activity" patented technology

This invention belongs to the fields of plant genetic engineering and biotechnology, and discloses a peanut endogenous strong promoter P. AhEF1α Its preparation methods and applications. Promoters include P AhEF1α‑1 P AhEF1α‑2 P AhEF1α‑3 and P AhEF1α‑4 This invention uses the Arabidopsis EF1α gene amino acid sequence as a reference, identifies candidate peanut EF1α genes through homology search, screens high-expression genes by expression level analysis, and clones the aforementioned promoter from the peanut genome of Yuhua 9326 using PCR technology. A recombinant vector containing this promoter is constructed and transformed into peanut embryogenic callus tissue. Functional verification shows that the promoter can efficiently drive the expression of downstream genes in peanut callus. This invention solves the technical bottleneck of existing peanut gene editing technologies that rely on exogenous promoters and have poor adaptability, fills the gap in the application of strong endogenous promoters in peanuts, and can be used to construct a precise gene editing system for peanuts, promoting stress-resistant, high-quality, and grade-equal peanut varieties. It has significant practical value and industrialization prospects.

The present application is based on the principle of protein termination-restart translation of ribosome shift, and provides a cloning method which can be used for improving the expression activity of membrane protein. The method is to construct a start-termination degenerate codon on a protein expression vector, and to restart the protein expression by using the ribosome shift phenomenon after the accidental termination of protein expression, so as to significantly reduce the protein expression rate, provide sufficient time for the folding and anchoring of membrane protein, and significantly improve the cloning method of membrane protein expression activity.