title: Structural and functional characterization of the actin-1 gene promoter from the Antheraea pernyi (Lepidoptera: Saturniidae).
abstract: The Chinese oak silkworm, Antheraea pernyi, is an economically important insect of the Saturniidae family. In this study, genome walking was performed to obtain an A. pernyi actin promoter, which can be employed in transgenic or stable cell line expression systems. The putative promoter was analyzed by the online promoter analysis programs at the Berkeley Drosophila Genome Project and the Web Promoter Scan Service, which led to the recognition of several functional elements. With respect to these elements, a series of actin A1 promoter fragments with 5'-deletions were generated that were then used to construct different vectors expressing Green Fluorescent Protein (GFP). The plasmids were transfected into Sf9 cells and GFP expression was determined by observing GFP fluorescence in cells and by measuring GFP mRNA levels with real-time polymerase chain reaction. Sequence comparisons indicated that the sequence cloned from A. pernyi was the actin A1 promoter. The basic function of the promoter was verified by constructing expression vectors and observing GFP expression. In addition, real-time polymerase chain reaction revealed a strong inhibitory element may exist upstream of the TATA box, which downregulated gene expression. The actin A1 promoter is an ideal candidate for use in A. pernyi transgenic systems.
authors.affiliation: School of Life Science and Biotechnology, Dalian University of Technology, Dalian, Liaoning 116023, People's Republic of China School of Agriculture, Eastern Liaoning University, Dandong, Liaoning 118000, People's Republic of China.
authors.forename: Danmei
authors.initials: D
authors.lastname: Liu
authors.affiliation: School of Life Science and Biotechnology, Dalian University of Technology, Dalian, Liaoning 116023, People's Republic of China.
authors.forename: Peng
authors.initials: P
authors.lastname: Geng
authors.affiliation: School of Life Science and Biotechnology, Dalian University of Technology, Dalian, Liaoning 116023, People's Republic of China.
authors.forename: Xiran
authors.initials: X
authors.lastname: Jiang
authors.affiliation: School of Life Science and Biotechnology, Dalian University of Technology, Dalian, Liaoning 116023, People's Republic of China.
authors.forename: Lijia
authors.initials: L
authors.lastname: An
authors.affiliation: School of Life Science and Biotechnology, Dalian University of Technology, Dalian, Liaoning 116023, People's Republic of China biolwl@dlut.edu.cn.
authors.forename: Wenli
authors.initials: W
authors.lastname: Li
mesh_terms: D000199:Actins / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D019476:Insect Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D009036:Moths / Q000235:genetics*; D011401:Promoter Regions, Genetic; D017422:Sequence Analysis, DNA
chemical_list: D000199:Actins; D019476:Insect Proteins
keywords: Antheraea pernyi; actin; functional analysis; promoter