Expression vector for promoting exocrine expression of target protein in escherichia coli cells

• Main Authors: NIU FANG, ZHAO FURONG, WANG SHENGBIN, LIN ZIHUANG, WANG ZHIPENG, PENG YANGXUAN
• Format: Patent
• Language: Chinese; English
• Published: 08.08.2023
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; ORGANIC CHEMISTRY; PEPTIDES; PROCESSES USING MICROORGANISMS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

The invention discloses an expression vector for promoting the exocrine expression of a target protein in escherichia coli cells, which at least comprises: (1) a target protein coding sequence and a YebF protein coding sequence, which are connected through a TEV enzyme digestion recognition site coding sequence; and (2) protein coding genes of DsbA, DsbC and SecM. According to the characteristics of secretory expression of escherichia coli, a YebF fusion tag is designed and can be used for guiding target protein to cross a plasma membrane and an outer membrane to achieve extracellular expression, meanwhile, an amino acid recognition site of TEV enzyme is arranged at the C-terminal of the YebF fusion tag, subsequent recovery and purification of the target protein are facilitated, Dsb family protein and SecM protein are co-expressed at the same time, and the expression efficiency is improved. The secretory expression quantity of the target protein can be obviously improved. 本发明公开了一种促进目标蛋白在大肠杆菌细胞外分泌表达的表达载体，至少包含：