Virus-Derived Construct Plasmid for Dual-Host Vector Production
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Solution Overview
Problem
Existing methods for producing virus-derived constructs require synthesis and construction of plasmids in Escherichia coli, limiting the versatility and efficiency of virus-derived construct production.
Innovation Solution
A method involving the use of a first and second microbial host cell to replicate and amplify a virus-derived construct plasmid, utilizing Bacillus subtilis and Escherichia coli, with specific nucleic acid sequences for replication and expression, enabling production in animal cells like human embryonic kidney cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If virus-derived construct plasmids are synthesized and constructed in Escherichia coli, then the construction process is established and reliable, but the versatility and efficiency of virus-derived construct production are limited
Solution Approach 1:
The patent creates a universal plasmid system that can function across multiple host organisms (E. coli, B. subtilis, and animal cells). The plasmid contains replication origins and regulatory elements that enable it to replicate and express genes in diverse hosts, thereby increasing the versatility of virus-derived construct production while maintaining a standardized construction approach.
Solution Approach 2:
The plasmid is designed with modular segments including distinct replication origins for different organisms, multiple selectable markers, and separated functional regions. This segmentation allows the plasmid to be adapted to different host systems while maintaining a relatively simple and established construction process in E. coli.
2Productivity
If virus-derived construct plasmids are produced in a combination of first and second microorganisms, then the productivity and genome copy numbers are enhanced, but the device complexity and process steps are increased
Solution Approach 1:
The plasmid is pre-designed and constructed in E. coli with all necessary replication origins, regulatory elements, and gene sequences incorporated before being transferred to the production host. This preliminary construction phase simplifies the overall process by preparing the plasmid in a well-established system before deployment in the dual-microorganism productivity enhancement strategy.
Solution Approach 2:
The plasmid itself acts as an intermediary carrier that bridges the two microorganism systems. It contains replication origins compatible with both first and second microorganisms, allowing it to replicate in both hosts and thereby enhance productivity without requiring complex inter-host communication or transfer mechanisms.
Data Source
AI summary
The present disclosure provides production of a virus-derived construct plasmid. In one aspect, the present disclosure provides a plasmid encoding at least some elements of a virus-derived construct. In one embodiment, the present disclosure provides a plasmid containing at least some of nucleic acid sequences required to construct a virus-derived construct in an operably linked form. In one embodiment, the nucleic acid sequence required to construct a virus-derived construct includes (A) a nucleic acid sequence including a terminal repeat, (B) a nucleic acid sequence encoding a packaging factor, (C) a nucleic acid sequence encoding a structural protein, and (D) a nucleic acid sequence encoding a functional cofactor.