METHOD FOR IMPROVING BIOCOUPLING EFFICIENCY BETWEEN PROTEIN AND NUCLEIC ACID BASED ON α-HELIX HANDLE

• Main Authors: LI, Xu, QU, Lihua, YU, Zhongbo, MA, Xiaofeng, HUANG, Wei
• Format: Patent
• Language: Chinese; English; French
• Published: 17.09.2020
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; ORGANIC CHEMISTRY; PEPTIDES; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

Provided is a method for improving the biocoupling efficiency between a protein and a nucleic acid based on an α-helix handle. The method comprises designing a handle H-tag carrying an unnatural amino acid, then constructing a fusion recombinant expression plasmid containing the H-tag and a protein to be tested, expressing and purifying a fusion recombinant protein containing the unnatural amino acid in the H-tag, and finally subjecting the unnatural amino acid in the H-tag of the fusion protein and a coupling group on a nucleic acid substrate to efficient connection by means of click chemistry. The method uses an α-helix handle to provide controllable protein surface reaction conditions for an unnatural amino acid, and avoids a complex structure, charges and polar nano environment on the surface of the protein to be tested, thereby achieving the target of the efficient connection of a protein and a nucleic acid by means of click chemistry. L'invention concerne un procédé pour améliorer l'efficacité de biocou