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

作     者：ZHU Yanbing YANG Feng 

作者机构：School of Biotechnology Jimei University Xiamen 361005 China Third Institute of Oceanography State Oceanic Administration Xiamen 361005 China 

出 版 物：《Acta Oceanologica Sinica》 (海洋学报（英文版）)

年 卷 期：2007年第26卷第2期

页      面：75-83页

核心收录：

学科分类：0710[理学-生物学] 0908[农学-水产] 0707[理学-海洋科学] 0906[农学-兽医学] 09[农学] 

基　　金：This research was supported by the National Natural Science Foundation of China under contract No.40276038 the"863"Program of China under contract No.2003AA626020 the Fujian Science Fundation under contract No.2003F001 

主　　题：white spot syndrome virus genome phage display library dot blot 

摘      要：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 × 10^*** 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.