Construction of prokaryotic expression vector of Peter Pan protein of Apis cerana
Qiu Huitong
Liu Yuming
Bai Hua
Cheng Yu
Li Ming
Fei Dongliang
Liu Yanxia
Ma Mingxiao
Abstract：This study aimed to investigate the role of Peter Pan(PPAN)in Chinese honeybee(Apis cerana)larvae.A prokaryotic expression plasmid was constructed to induce PPAN protein expression.The PPAN gene was amplified using PCR,and the amplified product was inserted into the prokaryotic expression vector pGEX-4T-1.The recombinant plasmid pGEX-4T-1-PPAN was successfully constructed and transformed into Escherichia coli BL21(DE3)competent cells.Optimal induction conditions for the target protein were analyzed via SDS-PAGE,and Western blot was performed for verification.Results demonstrated that PPAN protein was successfully expressed in inclusion body form in Escherichia coli.The optimal induction conditions were determined as follows:Induction temperature 28℃,induction time 8 h,and isopropyl β-D-1-thiogalactopyranoside(IPTG)concentration of 0.5 mmol/L.The amplified PPAN gene fragment measured 1 146 bp,encoding a protein with a molecular weight of 70.6 kDa.This study successfully established a prokaryotic expression system for PPAN protein in Escherichia coli,laying a foundation for further investigation into the impact of PPAN protein on Chinese Sacbrood Virus(CSBV)replication.
Keywords：Apis cerana larvaProkaryotic expressionPeter Pan(PPAN)
Publication Date：2025-06-15
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 7-11 )