Novel penaeus monodon ALFpm12 antibacterial peptide, and preparation method and applications thereof

Abstract

The invention belongs to the technical field of shelled animal gene engineering, and discloses a novel penaeus monodon ALFpm12 antibacterial peptide, and a preparation method and applications thereof.The novel penaeus monodon ALFpm12 antibacterial peptide is composed of 123 amino acids, the molecular weight is 13.54kDa, and the signal peptide is composed of 25 amino acids. The highest homology ofALFpm12 with disclosed penaeus monodon ALF antibacterial peptide is only 50%, up-regulating in organs of penaeus monodon infected by vibrio parahaemolyticus is realized, and the expression peak valueis achieved in 4h; the gene sequence can be obtained through RT-PCR of total RNA extracted from penaeus monodon tissues. The novel penaeus monodon ALFpm12 antibacterial peptide possesses inhibiting and killing effect on Staphylococcus aureus and Exiguobacterium Collins; the sterilization efficiency is high; the using amount is low; effect lasts long; production is simple; bacteria in sea water can be killed; and the application prospect in the field of aquaculture is promising.

Description

technical field [0001] The invention belongs to the technical field of crustacean genetic engineering, and in particular relates to a novel antibacterial peptide of Penaeus monodon ALFpm12, a preparation method and an application thereof. Background technique [0002] At present, the existing technologies commonly used in the industry are as follows: [0003] The ocean area accounts for two-thirds of the earth's surface, plays an important role in maintaining the global climate, and provides a stable ecological environment for the sustainable development of fisheries and aquaculture. Delicious, protein-rich fish, shrimp, and shellfish grow in the ocean. With the expansion and development of the marine aquaculture industry, by 2012, the aquaculture industry supplied 66,600 tons of seafood, including 175 species of marine invertebrates, mainly crustaceans and molluscs. Among them, farmed crustaceans accounted for 9.7% of the total volume of seafood, but accounted for 22.4% o...

AI Technical Summary

Problems solved by technology

However, with the development of intensive and semi-intensive aquaculture models in China and the increasing degree of intensification, eutrophic factors such as residual bait, feces, and aquatic animal and plant carcasses coexist in one water body, coupled with water pollution and The reduction of self-purification and regulation ability of ponds has led to severe excess of chemical oxygen demand, biological oxygen demand, ammonia nitrogen, nitrite, sulfide and other indicators in the aquaculture water, the increase in the types and numbers of pathogenic microorganisms, and the deterioration of the aquaculture environment has led to frequent fish and shrimp diseases occur

For example, in 2015, the success rate of vannamei farming in the Pearl River Delta was less than 20%

At present, the types and quantities of published shrimp antimicrobial peptides are far less than other species, which prevents their development and application

[0007] (2) The content of antimicrobial peptides in prawns is low, the extraction is difficult, and the cost is high

The technology of obtaining antimicrobial peptides through chemical synthesis also faces problems such as high cost and inferior activity to natural antimicrobial peptides.

None of the above methods is suitable for large-scale production and application of antimicrobial peptides

[0008] (3) The use of genetic engineering to produce antimicrobial peptides has the advantages of low cost and good industrialization prospects, but it must also solve the problems of easy formation of inclusion bodies, low activity, low yield of eukaryotic expression, and excessive modification brought about by prokaryotic expression.

[0009] Based on the above reasons, no shrimp antimicrobial peptides with high bactericidal activity and applicable to the prevention and treatment of shrimp diseases have been obtained so far.

Images