title: Characterization of actin and tubulin promoters from two sap-sucking pests, Nilaparvata lugens (Stål) and Nephotettix cincticeps (Uhler).
abstract: The brown planthopper, Nilaparvata lugens (N. lugens, Hemiptera: Delphacidae), and the green rice leafhopper, Nephotettix cincticeps (N. cincticeps, Hemiptera: Cicadellidae), two sap-sucking feeders, have caused many destructive agricultural disasters in East Asia, as they can bring diseases like 'hopper burn' and transmit plant viruses. Recently, continuously-cultured cell lines from both insects have been reported. However, exogenous protein expression systems have not yet been established. Here, we identified thirteen tubulin genes and three actin genes from N. lugens, and one tubulin gene and two actin genes from N. cincticeps. Furthermore, putative promoter regions of these genes were analyzed by bioinformatic approaches and 5'-RACE assay, and the promoter strength was evaluated by driving the enhanced green fluorescent protein expression in three insect cell lines, S2, Sf9, and BmN. Finally, we identified three effective promoters (Nl_αTub1 promoter, Nl_act3 promoter, and Nc_act1 promoter) among all candidates we screened. The Nc_act1 promoter showed the strongest activity, while the Nl_αTub1 promoter only worked in S2 cells. In conclusion, we identified and functionally characterized three native promoters from N. lugens and N.cincticeps, which would facilitate the establishment of exogenous protein expression systems suitable for these two insect pests.
authors.affiliation: College of Life Sciences, Jiangsu Normal University, Xuzhou 221116, China; Key Laboratory of Cell Proliferation and Differentiation of the Ministry of Education, State Key Laboratory of Membrane Biology, Department of Cell Biology and Genetics, College of Life Sciences, Peking University, Beijing 100871, China.
authors.forename: Nannan
authors.initials: N
authors.lastname: Qian
authors.affiliation: Key Laboratory of Cell Proliferation and Differentiation of the Ministry of Education, State Key Laboratory of Membrane Biology, Department of Cell Biology and Genetics, College of Life Sciences, Peking University, Beijing 100871, China.
authors.forename: Pengli
authors.initials: P
authors.lastname: Zheng
authors.affiliation: State Key Laboratory of Protein and Plant Gene Research, College of Life Sciences, Peking University, Beijing 100871, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Wang
authors.affiliation: College of Life Sciences, Jiangsu Normal University, Xuzhou 221116, China.
authors.forename: Shenyuan
authors.initials: S
authors.lastname: Pan
authors.affiliation: State Key Laboratory of Protein and Plant Gene Research, College of Life Sciences, Peking University, Beijing 100871, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Li
authors.affiliation: State Key Laboratory of Rice Biology and Ministry of Agriculture, Key Laboratory of Agricultural Entomology, Institute of Insect Sciences, Zhejiang University, Hangzhou 310058, China.
authors.forename: Chuanxi
authors.initials: C
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Cell Proliferation and Differentiation of the Ministry of Education, State Key Laboratory of Membrane Biology, Department of Cell Biology and Genetics, College of Life Sciences, Peking University, Beijing 100871, China; Center for Quantitative Biology, Peking University, Beijing 100871, China.
authors.forename: Jianguo
authors.initials: J
authors.lastname: Chen
authors.affiliation: Key Laboratory of Cell Proliferation and Differentiation of the Ministry of Education, State Key Laboratory of Membrane Biology, Department of Cell Biology and Genetics, College of Life Sciences, Peking University, Beijing 100871, China. Electronic address: junlinteng@pku.edu.cn.
authors.forename: Junlin
authors.initials: J
authors.lastname: Teng
mesh_terms: D000199:Actins / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D006430:Hemiptera / Q000145:classification / Q000235:genetics*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics*; D013045:Species Specificity; D014404:Tubulin / Q000235:genetics*
chemical_list: D000199:Actins; D014404:Tubulin
keywords: Actin; Insect cell line; Nephotettix cincticeps; Nilaparvata lugens; Promoter; Tubulin