cj2.2 Promoter Enhances Gene Expression Yield

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Solution Overview

Problem

Current methods for producing target products using microorganisms like Escherichia and Corynebacterium lack efficiency in promoting high yields of target genes, despite advances in gene expression systems and promoter research.

Innovation Solution

A novel promoter, cj2.2, is developed by modifying the existing cj1 promoter, exhibiting promoter activity greater than the cj1 promoter by about three times, and is integrated into a vector for expression in host cells to enhance gene expression levels and protein activity.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If conventional promoters (Ptac, Ptrc, Plac) derived from E. coli are used, then gene expression can be induced, but the expression efficiency and target product yield are insufficient

Engineering Contradiction:

Improvetarget product yieldVSAvoidexpression efficiency

Core Design Contradiction:

ProductivityVSReliability

Solution Approach 1:

The patent modifies the promoter sequence by changing specific nucleotide positions (e.g., positions -35 and -10 regions) to optimize binding affinity and transcription initiation efficiency, resulting in enhanced expression levels and target product yield

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The patent creates a composite promoter structure by combining functional elements from different promoter regions and optimizing the arrangement of regulatory sequences to achieve synergistic enhancement of transcriptional activity

Inventive Principle:

Principle #40Composite materials

2Productivity

If the cj1 promoter is used, then strong promoter activity (296% of tac promoter) is achieved, but further improvement in expression level and target product production efficiency is needed

Engineering Contradiction:

Improvetarget product production efficiencyVSAvoidpromoter sequence complexity

Core Design Contradiction:

ProductivityVSDevice complexity

Solution Approach 1:

The patent systematically modifies specific parameters of the cj1 promoter sequence, including nucleotide composition at key positions, spacing between regulatory elements, and consensus sequence matching, to progressively enhance promoter activity beyond the original cj1 level

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The patent focuses optimization on specific local regions of the promoter (such as the -35 box, -10 box, and spacer regions) rather than uniform modification throughout the entire sequence, achieving enhanced activity through targeted improvements in critical functional domains

Inventive Principle:

Principle #3Local quality

Data Source

PatentUS11603534B2Promoter and use thereof

Publication Date: 2023.03.14 CJ CHEILJEDANG CORP

AI summary

Provided are promoters and a method of producing L-amino acid using the same.