Transfection method for expressing freshwater crayfish cell exogenous gene and application thereof

Abstract

The invention relates to a transfection method for expressing a freshwater crayfish cell exogenous gene and application thereof, which relates to the biological technology in the aquatic animal field.The invention provides a strong promoter sequence for expressing the freshwater crayfish hemopoietic tissue cell exogenous gene, an expression vector suitable for expressing the freshwater crayfish hemopoietic tissue exogenous gene, an electroporation system suitable for expressing the freshwater crayfish hemopoietic tissue cell exogenous gene as well as a specific operation parameter and a transfection method suitable for expressing the freshwater crayfish hemopoietic tissue cell exogeneous gene. A recombinant plasmid of an extremely-early gene wsv249 promoter sequence comprising a white spot syndrome virus and a CeletrixTM electroporation system are used for studying the operation procedures and technical parameters for transfecting the freshwater crayfish hemopoietic tissue cell so asto establish a set of operation parameters and method for successively expressing the freshwater crayfish hemopoietic tissue cell exogenous gene.

Description

technical field [0001] The invention relates to biotechnology in the field of aquatic zoology, in particular to a transfection method for exogenous gene expression in freshwater crayfish cells and its application. Background technique [0002] my country is a big aquaculture country, and the shrimp farming industry (including seawater prawns and freshwater crayfish) is an important economic industry in many areas of my country. However, in recent years, disease problems, including bacterial diseases, fungal diseases, and viral diseases, have continued to be prominent, making the success rate of seawater shrimp farming low. In 2015, the success rate of shrimp farming in China was less than 10%, and the overall industry was in a downturn. Compared with seawater prawns, freshwater crayfish is an exotic species and has relatively low requirements for the breeding environment. However, in recent years, due to the large expansion of the breeding area and the continuous increase of...

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Problems solved by technology

For this reason, scientists from China, the United States, Sweden, Japan, Thailand, Vietnam and other countries have carried out extensive research on the immune defense mechanism of shrimp and the invasion mechanism of diseases, but the information obtained is still very limited, which seriously affects the production practice. guidance

The reason is largely due to the lack of a research platform for the transfection and expression of related exogenous genes, so that some key issues cannot be explored in depth.

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