Schizosaccharomyces Expression Vector Using ihc2 Terminator
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Solution Overview
Problem
Current expression systems using yeasts of the genus Schizosaccharomyces, such as S. pombe, face challenges in achieving high expression efficiency for foreign proteins, particularly due to the limitations of conventional terminators like nmt1, which affect the release of mRNAs and subsequent protein production.
Innovation Solution
Incorporating the ihc2 gene terminator from Schizosaccharomyces into expression vectors, which enhances the expression efficiency by optimizing mRNA release and protein production, outperforming conventional terminators like nmt1 and others.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional terminators (nmt1, SV40, hLPI) are used in expression vectors, then the vector structure is simple and easy to construct, but protein expression efficiency is low due to poor mRNA release
Solution Approach 1:
The patent changes the terminator parameter from conventional options (nmt1, SV40, hLPI) to the ihc2 terminator, which has superior mRNA release properties. This parameter change in the vector structure directly improves protein expression efficiency by enhancing transcription termination and mRNA processing, resolving the contradiction between simple vector design and high expression efficiency.
2Productivity
If terminator is not included in expression cassette, then vector construction is simpler, but mRNA release is insufficient and expression efficiency is low
Solution Approach 1:
The patent incorporates a terminator sequence (ihc2) that serves its function efficiently and then allows the system to proceed without requiring complex additional components. The terminator acts as a disposable functional element that completes the transcription process and enables high-efficiency expression without needing further structural modifications.
3Productivity
If various conventional terminators are tested to find optimal one, then expression efficiency can be improved, but time and resources are consumed
Solution Approach 1:
The patent performs preliminary identification and selection of the ihc2 terminator as the optimal element for Schizosaccharomyces pombe expression systems. By pre-determining the superior terminator based on its mRNA release properties and testing results showing at least 4 times higher expression than conventional terminators, the patent eliminates the need for time-consuming iterative testing in future applications.
Data Source
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AI summary
The object is to provide an expression vector capable of expressing with a higher expression efficiency a protein derived from a foreign structural gene by genetic engineering using a yeast of the genus Schizosaccharomyces as a host, a cloning vector to produce the expression vector, a method for producing the expression vector, a transformant containing an expression cassette of the expression vector, a method for producing the transformant, and a method for producing a protein using the transformant. A cloning vector comprising a promoter capable of functioning in a yeast of the genus Schizosaccharomyces, a cloning site for introducing a foreign structural gene which is located downstream from the promoter and is regulated by the promoter, and ihc2 gene terminator of the yeast of the genus Schizosaccharomyces, and an expression vector comprising the promoter, the foreign structural gene and the ihc2 gene terminator.