T7 expression system based on cytoplasm line plasmids and method for expressing protein in yeast by T7 expression system

Abstract

The invention relates to a T7 expression system based on cytoplasmic line plasmids, the T7 expression system comprises T7 RNA polymerase, a T7 transcription unit and cytoplasmic line plasmids, the T7 transcription unit is composed of a T7 promoter, a T7 terminator and a target gene, the system can be used for continuously, stably and efficiently expressing protein in yeast, and the line plasmids are from eukaryotic cells.

Description

technical field [0001] The present invention relates to a T7 expression system based on a cytoplasmic linear plasmid. The T7 expression system comprises T7 RNA polymerase, a T7 transcription unit and a cytoplasmic linear plasmid. The system can be used for continuous, stable and efficient protein expression in yeast. Background technique [0002] The T7 system comes from Escherichia coli T7 phage, which is a very powerful transcription system. The system is mainly composed of T7 RNA polymerase and its specifically recognized transcription unit initiated by the T7 promoter. At present, the application of the T7 expression system in prokaryotes is very mature, but the construction of a T7 expression system that can express proteins continuously, stably and efficiently in eukaryotes, especially Saccharomyces cerevisiae, is still limited, mainly because eukaryotes Nuclear transcribed mRNA faces the problem of nuclear export, and its mRNA has structural specificity. [0003] The...

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Problems solved by technology

[0006] For eukaryotes, some researchers have constructed a nuclear genome-based T7 expression system in Saccharomyces cerevisiae. However, due to the lack of a 5′ cap structure, a large amount of T7 mRNA in the nucleus cannot enter the cytoplasm, and it binds to ribosomes and is translated into protein. The T7 expression system did not express the target protein (see Non-Patent Document 3)

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