title: Enhanced green fluorescent protein (egfp) gene expression in Tetraselmis subcordiformis chloroplast with endogenous regulators.
abstract: On the basis of fundamental genetic transformation technologies, the goal of this study was to optimize Tetraselmis subcordiformis chloroplast transformation through the use of endogenous regulators. The genes rrn16S, rbcL, psbA, and psbC are commonly highly expressed in chloroplasts, and the regulators of these genes are often used in chloroplast transformation. For lack of a known chloroplast genome sequence, the genome-walking method was used here to obtain full sequences of T. subcordiformis endogenous regulators. The resulting regulators, including three promoters, two terminators, and a ribosome combination sequence, were inserted into the previously constructed plasmid pPSC-R, with the egfp gene included as a reporter gene, and five chloroplast expression vectors prepared. These vectors were successfully transformed into T. subcordiformis by particle bombardment and the efficiency of each vector tested by assessing EGFP fluorescence via microscopy. The results showed that these vectors exhibited higher efficiency than the former vector pPSC-G carrying exogenous regulators, and the vector pRFA with Prrn, psbA-5'RE, and TpsbA showed the highest efficiency. This research provides a set of effective endogenous regulators for T. subcordiformis and will facilitate future fundamental studies of this alga.
authors.affiliation: Key Laboratory of Coastal Biology and Biological Resource Utilization, Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Chunhui Road 17, Laishan District, Yantai City, 264003, Shandong Province, China.
authors.forename: Yulin
authors.initials: Y
authors.lastname: Cui
authors.affiliation: Key Laboratory of Coastal Biology and Biological Resource Utilization, Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Chunhui Road 17, Laishan District, Yantai City, 264003, Shandong Province, China.|University of Chinese Academy of Sciences, Yuquan Road 19, Shijingshan District, Beijing City, 100049, China.
authors.forename: Jialin
authors.initials: J
authors.lastname: Zhao
authors.affiliation: College of Life Sciences, Yantai University, Qingquan Road 30, Laishan District, Yantai City, 264005, Shandong Province, China.
authors.forename: Shichang
authors.initials: S
authors.lastname: Hou
authors.affiliation: Key Laboratory of Coastal Biology and Biological Resource Utilization, Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Chunhui Road 17, Laishan District, Yantai City, 264003, Shandong Province, China. sqin@yic.ac.cn.
authors.forename: Song
authors.initials: S
authors.lastname: Qin
mesh_terms: D000460:Chlorophyta / Q000235:genetics* / Q000378:metabolism; D002736:Chloroplasts / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D014170:Transformation, Genetic
chemical_list: C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins
keywords: Bombardment; Chloroplast; Promoters; Tetraselmis subcordiformis