Method for constructing highly expressed trehalose synthase engineering bacteria by using Pcry3Aa promoter

A trehalose synthase and promoter technology, applied in the field of genetic engineering, can solve the problems of difficult manufacturing, complicated process and high extraction cost
CN105779489AActive Publication Date: 2016-07-20山东开盾生物科技有限公司
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
山东开盾生物科技有限公司
Publication Date
2016-07-20

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Abstract

The invention relates to a method for constructing highly expressed trehalose synthase engineering bacteria by using a Pcry3Aa promoter. For a recombinant carrier, a Pcry3Aa-PhoD fragment by which a Pcry3Aa promoter fragment and a PhoD signal peptide fragment are connected by an overlap PCR (Polymerase Chain Reaction) is inserted at the upstream of a restriction enzyme cutting site BamHI of a shuttle plasmid PHT01, and a target protein trehalose synthase TreS fragment is inserted between two restriction enzyme cutting site, i.e., BamHI and AatII. The invention further relates to a method for constructing the highly expressed trehalose synthase engineering bacteria by using the recombinant carrier. According to the method disclosed by the invention, the Pcry3Aa promoter is adopted to naturally induce the synthesis of trehalose synthase; because the Pcry3Aa promoter contains a special STAB-SD structure, the stability of the Pcry3Aa promoter to transcribe mRNA is improved, the half-life period of mRNA is prolonged, the mRNA translation level of a downstream target gene is improved, and therefore the trehalose synthase is highly expressed.
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Description

technical field

[0001] The invention relates to a method for constructing high-expression trehalose synthase engineering bacteria by using a Pcry3Aa promoter, which belongs to the technical field of genetic engineering. Background technique

[0002] Trehalose, also known as cocoon honey, mushroom sugar or mushroom sugar, is not reducing, and its molecular formula is C 12 h 22 o 11 2H 2 O. Trehalose can not only store energy, but also protect the body under special circumstances. For example, under harsh conditions such as high osmotic pressure, high temperature, and dryness, trehalose can protect organisms against external adverse factors and maintain the stability of organisms. Trehalose can only be synthesized in harsh environments, and will be quickly degraded in suitable environments, which brings difficulties to its production, which also leads to its high price. Therefore, finding a way to synthesize trehalose in large quantities is a technical problem that needs...

Claims

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