Eukaryotic expression of chitinase gene from Lymantria dispar and construction of recombinant baculovirus

Author of the article:FAN Xiao-Jun , SONG Zhi-Fang, XIAN Xiao-Xiao,REN H

Author's Workplace：College of Chemical Engineering and Technology, Taiyuan University of Technology

Key Words： Biological Control; Lymantria dispar; gene clone; chitinase; recombinant baculovirus

Abstract：      Chitin is an important part of exoskeletons and peritrophic membrane of insect. In view of chitinase playing a pivotal role in the process of insect development, application of insect chitinase provides a strategy for exploring new methods for biological control of pests. In this paper, based on Autographa californica nuclear polyhedrosis virus polyhedrin gene sequence and the open reading frame of I chitinase gene from Lymantria dispar couples of primers were designed to clone the above two genes, with total length of 783 bp and 1 737 bp, respectively. Recombinant plasmid pFastBac-LdCht and pFastBac-AcPH-LdCht were constructed and transformed into E. coli DH10Bac to form recombinant bacmids which then were transfected into Sf9 cells to produce recombinant baculovirus AcMNPV-AcPH--LdCht and AcMNPV-LdCht. They were used to express the protein and get recombinant virus respectively. Cells successfully expressed active chitinase and amplified recombinant viruses in the Helicoverpa armigera. This study provided the basis for in-depth understanding of the insect chitinase and a potential way for recombinant virus application.