Random Diversification of Bacteriophage Tail Fibres
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
The development of bacteriophage cocktails for treating bacterial infections is hindered by the need for individual selection and testing of bacteriophages, which is time-consuming and limited to specific bacterial species, and the challenge of modifying bacteriophages to target a broader range of hosts while maintaining infectivity.
Innovation Solution
A method for random diversification of nucleotide sequences encoding target proteins of bacteriophages using PCR, preserving the identity of certain domains, to create recombinant bacteriophages with variable host specificity, allowing for the production of bacteriophages with extended infectivity spectra.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If individual selection and testing of bacteriophages is performed, then reliability of treatment is improved, but time required for development increases
Solution Approach 1:
The patent segments the bacteriophage population into multiple variants with different host specificities by diversifying specific domains of the tail fibre proteins. This allows parallel evaluation of multiple phage types simultaneously rather than sequential testing, reducing development time while maintaining treatment reliability through the diversified phage bank.
Solution Approach 2:
The patent changes the genetic parameters of bacteriophages by introducing random mutations in specific domains of the tail fibre proteins while maintaining other regions. This creates a diverse bank of phage variants with different host ranges, enabling reliable treatment without time-consuming individual selection.
2Adaptability or versatility
If bacteriophages are modified to target broader host ranges, then adaptability is improved, but loss of specificity for particular bacterial species increases
Solution Approach 1:
The patent divides the tail fibre protein into distinct functional domains: a conserved core region maintaining structural integrity and a variable domain responsible for host recognition. By mutating only the variable domain while preserving the conserved core, the method achieves broad host range adaptability without completely losing specificity for particular bacterial species.
Solution Approach 2:
The patent applies local quality by making different parts of the tail fibre protein have different properties: the conserved core maintains stable structural characteristics for phage assembly, while the variable domain provides diverse host recognition capabilities. This localized differentiation enables simultaneous achievement of broad adaptability and maintained specificity.
3Quantity of substance
If random diversification of nucleotide sequences is performed, then quantity of diverse bacteriophage variants increases, but manufacturing complexity of the process increases
Solution Approach 1:
The patent applies local quality by targeting random diversification to specific variable domains while preserving conserved regions. This localized approach generates diverse variants efficiently without requiring complex genome-wide mutagenesis protocols, thereby increasing the number of variants while controlling process complexity.
Solution Approach 2:
The patent performs preliminary action by designing primers that specifically target the conserved regions of the tail fibre proteins. These primers serve as anchors for PCR amplification, enabling systematic diversification of variable domains through a standardized protocol that scales well to produce large numbers of variants without proportionally increasing complexity.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach enables the generation of a diverse bank of recombinant bacteriophages capable of infecting a wider range of hosts, overcoming the limitations of traditional methods by increasing the spectrum of activity and adaptability of bacteriophages, thus providing a more universal treatment option for bacterial infections.
Implementation Method 1
an error-prone PCR is performed on the whole of sequence S using primers corresponding to segments F1 and F2, by which sequence S will be mutated randomly on its entire length
Implementation Method 2
error-prone PCR is performed on the whole of sequence S using primers corresponding to segments F1 and F2, by which sequence S will be mutated randomly
Implementation Method 3
high-fidelity PCR is carried out starting from the amplification products eluted in stage ii), using the primer pairs corresponding respectively to at least F1 and D, and F2 and D, in order to amplify the F1-D and D-F2 regions of S mutated in stage i) whose inner segment D has preserved its identity
Data Source
AI summary
The invention relates to a very general method for the random diversification of a nucleotide sequence S by PCR while preserving the identity of some domains of said sequence S; the invention also relates to a bank of nucleotide sequence thus diversified, and to diversified proteins obtained by the expressions of the nucleotide sequences in an appropriate host.