Construction of white spot syndrome virus (WSSV) whole genome phage display library

A rebuilt vector pCANTAB 5 EE was obtained by inserting a 34 bp double-stranded oligonucleotide which contained a EcoRV recognition sequence into pCANTAB 5 E.White spot syndrome virus (WSSV) genome DNA was fragmented by sonication to isolate fragments mainly in the range of 0.8~2.0 kb, then the fragments were blunt-ended with T4 DNA polymerase and cloned into the EcoRV site of pCANTAB 5 EE.The primary recombinant clone of the library was 3.0×10⁵.Colony PCR of random selected recombinants showed that the size of the inserts was 0.12~1.77 kb.After the whole library recombinant phages infected Escherichia coli HB2151 cells, the extracellular and periplasmic extracts were dropped on PVDF membranes to perform dot blot, using polyclonal mouse anti-VP24 serum, anti-WSV026 serum, anti-WSV063 serum, anti-WSV069 serum, anti-WSV112 serum, anti-WSV238 serum, anti-WSV303 serum and anti-VP26 serum as the primary antibody, respectively.The results showed that the display library could express the viral proteins.