Abstract: The present invention relates to a pseudotyped insect baculovirus gene transfer system and a virus for shrimps, and a construction method and use thereof. The present invention belongs to the technical field of genetic engineering. The pseudotyped insect baculovirus gene transfer system for shrimps disclosed in the present invention includes a Bac-to-Bac insect baculovirus expression system, an expression plasmid carrying shrimp virus envelope protein gene and an insect packaging cell. The present invention can achieve stable and efficient transfer and expression of a foreign gene in a shrimp tissue and cell.

Abstract: By establishing effective methods for shrimp 3D cell culture and passage, the present invention provides a technology of continuous shrimp cell culture intended for the establishment of immortalized shrimp cell lines. The present invention provides a preparation method of matrigel for 3D cell culture of shrimp by optimizing an additive proportion of matrigel. The present invention further provides a technology of separation and 3D cell culture of shrimp haemolymph cells, where shrimp haemolymph cells adhere to and grow on the surface of the matrigel in the form of a single round cell and a cell pellet/cellular spheroid, with survival and growth abilities being superior to 2D culture effects. The above technology is achieved by optimizing a formula of complete medium for shrimp cells, selecting the medium as an anticoagulant and a diluent for shrimp haemolymph cells, selecting a 3D culture method for surface-adhered growth in the matrigel.

Abstract: The present invention relates to a pseudotyped insect baculovirus gene transfer system and a virus for shrimps, and a construction method and use thereof. The present invention belongs to the technical field of genetic engineering. The pseudotyped insect baculovirus gene transfer system for shrimps disclosed in the present invention includes a Bac-to-Bac insect baculovirus expression system, an expression plasmid carrying shrimp virus envelope protein gene and an insect packaging cell. The present invention can achieve stable and efficient transfer and expression of a foreign gene in a shrimp tissue and cell.

Abstract: By establishing effective methods for shrimp 3D cell culture and passage, the present invention provides a technology of continuous shrimp cell culture intended for the establishment of immortalized shrimp cell lines. The present invention provides a preparation method of matrigel for 3D cell culture of shrimp by optimizing an additive proportion of matrigel. The present invention further provides a technology of separation and 3D cell culture of shrimp haemolymph cells, where shrimp haemolymph cells adhere to and grow on the surface of the matrigel in the form of a single round cell and a cell pellet/cellular spheroid, with survival and growth abilities being superior to 2D culture effects. The above technology is achieved by optimizing a formula of complete medium for shrimp cells, selecting the medium as an anticoagulant and a diluent for shrimp haemolymph cells, selecting a 3D culture method for surface-adhered growth in the matrigel.