Prokaryotic expression of Nilaparvata lugens flightin gene and its differential expression in the insect

Author of the article:Xiao-Qin Zhang, Yu-Qiong Hu，Chuan-Xi Zhang

Author's Workplace：Institute of Insect Science, Zhejiang University

Key Words：the brown planthopper, flightin, prokaryotic expression, polyclonal antibody, wing dimorphism，developmental expression

Abstract：Flightin, located in thick filament, was originally found in indirect flight muscle of Drosophila melanogaster. It is important for structural integrity and sarcomere function. The difference of the muscle components between macropterous and brachypterous adults of the brown planthopper (BPH, Nilaparvata lugens) was unknown yet. We amplified flightin gene from cDNA of macropterous female adults and inserted it into the expression vector pGEX-6P-1 for fusion expression with gultathione S transferase (GST).The recombinant vector was then tranformed into Escherichia coli Rosseta. After optimization of the expression conditions of different temperatures and concentrations of IPTG, high level expression of flightin was archieved. The soluble expressed GST-flightin was purified and was further used to immune rabbit for preparing the polyclonal antibody against flightin. Using the prepared antibody, we detected that flightin expression in different developmental stages and in two wing forms. The results showed that flightin was uniquely expressed in adult stage and only expressed in macropterous female adults, no positive bands were detected in eggs, nymphs and brachypterous female adults. This study laid a basis for the study of wing muscle development and wing dimorphism，as well as for research of the interaction of flightin with other proteins in BPH.