| Construction of white spot syndrome virus (WSSV) whole genome phage display library |
| |
| 作者姓名： | ZHU Yanbing YANG Feng |
| |
| 作者单位： | ZHU Yanbing1，YANG Feng21. School of Biotechnology，Jimei University，Xiamen 361005，China2. Third Institute of Oceanography，State Oceanic Administration，Xiamen 361005，China |
| |
| 基金项目： | 国家自然科学基金
,
国家高技术研究发展计划(863计划)
,
the Fujian Science Fundation |
| |
| 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 3.0×105. 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.
|
| 关 键 词： | 对虾 白斑综合症病毒 全基因组 噬菌体展示文库 WSSV |
| 收稿时间： | 2006-03-09 |
| 修稿时间： | 2006-07-20 |
Construction of white spot syndrome virus (WSSV) whole genome phage display library |
| |
| ZHU Yanbing,YANG Feng.Construction of white spot syndrome virus (WSSV) whole genome phage display library[J].Acta Oceanologica Sinica,2007,26(2):75-83. |
| |
| Authors: | ZHU Yanbing and YANG Feng |
| |
| Institution: | 1.School of Biotechnology, Jimei University, Xiamen 361005, China2.Third Institute of Oceanography, State Oceanic Administration, Xiamen 361005, China |
| |
| Abstract: | 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. |
| |
| Keywords: | white spot syndrome virus genome phage display library dot blot |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
| 点击此处可从《海洋学报(英文版)》浏览原始摘要信息 |
| 点击此处可从《海洋学报(英文版)》下载免费的PDF全文 |
|
