Pet28a expression vector

A polyclonal antibody against P. fucata IRR was made by Shanghai Biomodel Organism Science & Technology Development Co., Ltd (Shanghai, China). An 837-bp cDNA fragment encoding the TK domain of IRR (GenBank accession number: JX121113) was amplified and cloned into the BamHI/XhoI sites of the pET28a expression vector (TAKARA, Japan), and histidine-tagged TK was expressed in E. coli and purified with a Ni²⁺-NTA column.
Polyclonal antibody was raised against the purified TK domain in two rabbits. After immunization, the antisera were collected and purified by a protein A affinity column. The titers of the antisera were determined by ELISA. Western blot analyses were conducted to characterize the polyclonal antibody.

The 95% abamectin was ordered from Bangnong Chemical Company (Guangzhou, China), 5‐nitrouracil was ordered from Adamas‐beta (Shanghai, China); iQ™ SYBR^® Green Supermix was acquired from Bio‐Rad (Hercules, CA, USA); α‐naphthol was ordered from Shanghai Chemical Reagent Company of the Chinese Medical Group (Shanghai, China); pGEM‐T Easy Vector was ordered from Promega (Madison, WI, USA); pET‐28a expression vector and EcoR I, Nde I were obtained from Takara (Dalian, China). Trans5α and BL21 (DE3) competent cells were acquired from TransGen Biotech (Beijing, China); isopropyl b‐D‐1‐thiogalactopyranoside (IPTG), UDP‐glucose and kanamycin were obtained from Sigma Chemical Co. (St Louis, MO, USA). Other chemicals and reagents were high‐quality commercially available products supplied by local suppliers.