Transfection of crayfish hematopoietic tissue cells

Shi, HongState Ocean Adm, State Key Lab Breeding Base Marine Genet Resource, Key Lab Marine Genet Resources,Inst Oceanog 3, Fujian Key Lab Marine Genet Resources,South China, Xiamen 361005, Peoples R China (författare)

Transfection is a powerful tool useful for studying gene function. Establishing transfection methods that enable highly efficient DNA uptake has become increasingly important. The crayfish hematopoietic tissue (Hpt) cell cultures have been proven to be suitable for studies on immunity and cell differentiation in crustaceans including shrimps, but no efficient gene transfer and expression method is available for these cells. Here we report a novel and highly efficient DNA transfection system based on electroporation. This method depends on a recombinant plasmid with the promoter from white spot syndrome virus immediate-early gene wsv249. This plasmid could be introduced into primary cells and efficiently express foreign genes by electroporation. By optimizing different electroporation parameters, more than 30% transfection efficiency could be achieved with the relative viability of cells around 50%. This is the first report of gene introduction to crayfish Hpt cells and will be useful for the expanding our research on crustacean immunity.

Ruan, LingweiState Ocean Adm, State Key Lab Breeding Base Marine Genet Resource, Key Lab Marine Genet Resources,Inst Oceanog 3, Fujian Key Lab Marine Genet Resources,South China, Xiamen 361005, Peoples R China (författare)
Söderhäll, IreneUppsala universitet,Jämförande fysiologi(Swepub:uu)irsoderha (författare)
Söderhäll, KennethUppsala universitet,Jämförande fysiologi(Swepub:uu)kennsode (författare)
Xu, XunState Ocean Adm, State Key Lab Breeding Base Marine Genet Resource, Key Lab Marine Genet Resources,Inst Oceanog 3, Fujian Key Lab Marine Genet Resources,South China, Xiamen 361005, Peoples R China (författare)
State Ocean Adm, State Key Lab Breeding Base Marine Genet Resource, Key Lab Marine Genet Resources,Inst Oceanog 3, Fujian Key Lab Marine Genet Resources,South China, Xiamen 361005, Peoples R ChinaJämförande fysiologi (creator_code:org_t)

Ingår i:Developmental and Comparative Immunology: Elsevier BV88, s. 70-760145-305X1879-0089

Av författaren/redakt...: Shi, Hong; Ruan, Lingwei; Söderhäll, Irene; Söderhäll, Kenne ...; Xu, Xun

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