Constructed carp apoptosis gene, vector and its application

Abstract

The invention relates to a constructed carp apoptotic gene, a vector and application of the gene. From 5' to 3', the constructed carp apoptotic gene is sequentially composed of carp MT-1 promoter, caspase-8 small subunit (P10) coding sequence, caspase-8 large subunit (P20) coding sequence, and carp MT-1polyA signal sequence. The carp DNA vaccine vector of the invention includes a eukaryotic expression vector and an apoptotic gene of the above carp gene in the eukaryotic expression vector. The inductive apoptotic gene is constructed based on carp MT promoter and caspase-8, and after being inserted to pEGFP-N1 vector to construct 'suicide' DNA vaccine vector, the apoptotic gene may respond to the induction of the conditions such as metal ions (e.g., Zn2+) to express apoptosis of caspase-8 induced transfection cells and may apply to the preparation of carp DNA vaccines to further improve safety of DNA vaccines.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a constructed carp apoptosis gene, a carrier and an application thereof. Background technique [0002] Since the advent of DNA vaccine, it has become an important direction of new vaccine research because of its advantages of simple preparation, transportation and storage, and the ability to induce humoral immunity and cellular immunity at the same time. However, the continuous expression of DNA after vaccination may induce the host's own immune response and immune tolerance, and increase the risk of vaccine DNA integrating into the host genome. Although the "suicide" DNA vaccine based on the alphavirus replicon greatly shortens the existence time of the vaccine DNA in the body, this type of vaccine vector also has some disadvantages: (1) the fragment of the alphavirus replicon is too large (> 7kb), plus the necessary elements such as plasmid replication and screening...

AI Technical Summary

Problems solved by technology

However, continuous expression after DNA vaccination may induce host autoimmune response and immune tolerance, and increase the risk of vaccine DNA integration into the host genome

Although the "suicide" DNA vaccine based on the alphavirus replicon greatly shortens the existence time of the vaccine DNA in the body, this type of vaccine vector also has some disadvantages: (1) the fragment of the alphavirus replicon is too large (> 7kb), plus the necessary elements such as plasmid replication and screening, reporter gene, etc., the vaccine vector exceeds 11kb, which affects the capacity of the vector, and because the DNA vaccine inserted with the foreign gene is too large, it also increases the difficulty of vaccine construction and affects Transfection efficiency

(2) The process of apoptosis is uncontrollable. After 2 to 5 days after inoculation of the "suicide" DNA vaccine, the vaccine will be eliminated. If the process of apoptosis can be delayed, it is possible to obtain a better immune effect

The continued expression of foreign genes after vaccination may induce host autoimmune responses and immune tolerance, and increase the risk of vaccine DNA integrating into the host genome

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