Microorganisms with Reduced Competence via comK Gene Mutation
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Solution Overview
Problem
Microbial strains, particularly probiotics used as feed additives, can acquire unwanted antibiotic resistance genes through their natural competence to take up DNA from the environment, leading to undesirable genetic changes.
Innovation Solution
Introducing point mutations into the comK gene, which encodes the competence transcription factor, to knock out the competence machinery in microorganisms, thereby reducing their ability to take up DNA by at least 50%, and in some cases, completely eliminating this capability.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If the competence machinery is left intact in microbial strains, then the strains can acquire new characteristics and adapt to the environment through natural transformation, but the strains may acquire unwanted antibiotic resistance genes which is not desirable
Solution Approach 1:
The invention extracts and removes the competence machinery from the microbial strain by knocking out the comK gene. This eliminates the ability to take up extracellular DNA while preserving other beneficial characteristics of the strain, directly resolving the contradiction between adaptability and harmful gene acquisition.
Solution Approach 2:
The invention converts the harmful capability of natural transformation (which can acquire unwanted genes) into a benefit by selectively removing only the DNA uptake function. The strain retains its probiotic properties and adaptability through other mechanisms while being protected from acquiring harmful antibiotic resistance genes.
2Object-affected harmful factors
If the comK gene is knocked out to reduce competence, then the ability to take up DNA is reduced by at least 50%, but there may be detrimental effects on the characteristics of the cells
Solution Approach 1:
The invention specifically extracts and removes only the comK transcription factor gene responsible for competence regulation. This targeted approach ensures that only the DNA uptake function is eliminated while other cellular functions and probiotic activities remain intact, thus maintaining reliability.
Solution Approach 2:
The invention applies local quality change by modifying only the specific comK gene locus while leaving the rest of the genome unchanged. This localized modification ensures that the reduction of harmful DNA uptake capability does not negatively impact other cellular characteristics and probiotic functions.
3Object-affected harmful factors
If point mutations are introduced into the comK gene to knock out competence, then the competence machinery is disabled, but the complexity of the genetic modification process increases
Solution Approach 1:
The invention changes the genetic parameter of the comK gene by introducing specific point mutations that create premature stop codons. This simple parameter change (nucleotide substitution) effectively knocks out the gene function with minimal complexity compared to larger deletions or insertions.
Solution Approach 2:
The invention extracts the functional capability of the comK gene by introducing stop codons that prevent translation. This approach disables the gene with simple point mutations rather than requiring complex large-scale genetic modifications, thus reducing process complexity.
Data Source
AI summary
The subject of the present invention are microorganisms with reduced competence and compositions containing such microorganisms, in particular feed, food and pharmaceutical compositions.