Construction of cytomegalovirus promoter-driven gene expression system in Laodelphax striatellus  

作　　者：Xiaohui Cheng Wan Zhao Guohua Liang Hong Lu Yumei Fu Yiming Li Feng Cui 

机构地区：[1]State Key Laboratory of Integrated Management of Pest Insects and Rodents,Insttute of Zoology,Chinese Academy of Sciences,Bejing,China [2]University of Chinese Academy of Sciences,Bejing,China [3]School of Life Sciences,Hebei University,Baoding,Hebei China [4]Key Laboratory of Animal Ecology and Conservation Biology,Institute of Zoology,Chinese Academy of Sciences,Bejing,China [5]Institute of Wetland Ecology&Clone Ecologyl Zhejiang Provincial Key Laboratory of Plant Evolutionary Ecology and Conservation,Taizhou University,Taizhou,Zhejiang,China

出　　处：《Insect Science》2024年第3期720-732,共13页昆虫科学（英文版）

基　　金：supported by grants from the National Natural Science Foundation of China(No.32272532);the Strategic Priority Research Program of the Chinese Academy of Sciences(No.XDA26050404);the National Natural Science Foundation of China(No.32230090);the Youth Innovation Promotion Association of CAS(No.2019086).

摘　　要：The small brown planthopper(SBPH,Laodelphax striatellus)is a significant rice pest,responsible for transmitting rice stripe virus(RSV)in a persistent and propagative manner.RSV is one of the most detrimental rice viruses,causing rice stripe disease,which results in considerable loss of rice grain yield.While RNA interference and gene knockout techniques have enabled gene downregulation in SBPH,no system currently exists for the overexpression of endogenous or exogenous genes.Consequently,the development of a protein expression system for SBPH is imperative to serve as a technical foundation for pest control and gene function investigations.This study aimed to construct an expression vector using the promoter of the constitutive-expressed tubulin gene of SBPH,and promoter of human cytomegalovirus(CMV).Fluorescence experiments demonstrated that both tubulin and CMV promoter could drive green fluorescent protein(GFP)expression in SBPH,and could also facilitate the expression of a nucleocapsid protein(NP)-GFP fusion protein containing viral NP with comparable efficiency.Through expression vector optimization,we have identified that the 3 tandem CMV promoters display a significantly higher promoter activity compared with both the 2 tandem CMV promoters and the single CMV promoter.In addition,the incorporation of Star polycation nanoparticles significantly enhanced the expression efficiency in SBPH.These results provide a promising technical platform for investigating gene functions in SBPH.

关 键 词：cytomegalovirus promoter gene expression system NANOPARTICLE small brown planthopper 

分 类 号：Q96[生物学—昆虫学]