Method for simultaneously cloning multiple exogenous genes to microbial genome

Abstract

The invention discloses a method for simultaneously cloning multiple exogenous genes to a microbial genome. The method comprises steps of introducing a plurality of exogenous genes to a plurality of bidirectional gene expression carriers, and then simultaneously introducing the plurality of bidirectional gene expression carriers into a host microbe. Each bidirectional gene expression carrier comprises a bidirectional terminator and a bidirectional promoter. Except for the first bidirectional gene expression carrier and the last bidirectional gene expression carrier, the 3' ends of the bidirectional terminators of other bidirectional gene expression carriers and the 5' ends of the bidirectional terminators of the next bidirectional gene expression carriers have the same homologous arm. The 5' end of the bidirectional terminator of the first bidirectional gene expression carrier and the 3' end of the bidirectional terminator of the last bidirectional gene expression carrier can both carry out homologous recombination with the genomes of a host microbe. Through the provided method, established is a novel technology that can organically integrate the whole gene expression process such as expression design, clone design, host cell modification, and the like.

Description

technical field [0001] The invention belongs to the field of genetic engineering and relates to a method for simultaneously cloning multiple exogenous genes into microbial genomes. Background technique [0002] The transformation of microbial DNA, including the knockout and overexpression of target genes, is the core content of the field of genetic engineering. The expression of one or two genes in the target microorganism is not difficult to achieve through existing technologies, however, there are still many difficulties in the expression of multiple genes (more than two) according to the existing technologies, such as long cloning cycle, heavy workload, and low success rate. Low, low upper limit of gene expression quantity, etc. Especially for gene expression in eukaryotes, generally each gene needs a separate promoter and terminator, so that one gene expression requires 3 cloning, and if multi-gene expression is performed, such as 6 gene expression, 18 clones are requir...

AI Technical Summary

Problems solved by technology

However, the vectors in these patents are generally constructed by the inventors with fixed promoters or other transcription elements, and can only express the target gene at a fixed intensity

However, the current genetic engineering transformation work often requires the coordination of multiple genes, that is, the expression intensity of each gene needs to be different according to the designer's intention, which is difficult to achieve with existing tool carriers or technologies, and this technology will solve this problem

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