title: Construction of Plastid Expression Vector and Development of Genetic Transformation System for the Seaweed Pyropia yezoensis.
abstract: Pyropia yezoensis, belonging to the Rhodophyta, is an economically important seaweed. In this study, we developed a high-efficiency plastid transformation platform for P. yezoensis. In the plastid transformation vector, psbA UTR of P. yezoensis, including the promoter and 3' UTR, was used to express foreign genes. The integration site was a transcriptionally active intergenic region between the rrsB and trnI genes, located in the inverted repeat regions of the plastid genome. The CAT and eGFP genes were integrated into the plastid genome at this site. The expression of CAT in the transformants confers resistance to chloramphenicol through the action of chloramphenicol acetyltransferase, which inactivates the drug, thereby allowing the plant to grow well under selective pressure. The eGFP fluorescence signal was also observed in transformed cells and the transformants. The average survival rate of treated cells was estimated to be approximately 4.2‰ (4 transplastomic colonies per 1000 gametophyte cells). Multiple-PCR analyses confirmed that the CAT and eGFP genes were successfully integrated in the site between rrsB and trnI. Western blot also showed eGFP expression in the cells of transformants. Thus, this study presents the first convenient plastid gene expression system for P. yezoensis and provides an important platform for studying gene function in P. yezoensis.
authors.affiliation: College of Marine Life Sciences, Ocean University of China, Qingdao, 266003, China. fnkong@ouc.edu.cn.|Key Laboratory of Marine Genetics and Breeding of Ministry of Education, Ocean University of China, Qingdao, 266003, China. fnkong@ouc.edu.cn.
authors.forename: Fanna
authors.initials: F
authors.lastname: Kong
authors.affiliation: College of Marine Life Sciences, Ocean University of China, Qingdao, 266003, China.|Key Laboratory of Marine Genetics and Breeding of Ministry of Education, Ocean University of China, Qingdao, 266003, China.
authors.forename: Hailong
authors.initials: H
authors.lastname: Zhao
authors.affiliation: College of Marine Life Sciences, Ocean University of China, Qingdao, 266003, China.|Key Laboratory of Marine Genetics and Breeding of Ministry of Education, Ocean University of China, Qingdao, 266003, China.
authors.forename: Weixun
authors.initials: W
authors.lastname: Liu
authors.affiliation: College of Marine Life Sciences, Ocean University of China, Qingdao, 266003, China.|Key Laboratory of Marine Genetics and Breeding of Ministry of Education, Ocean University of China, Qingdao, 266003, China.
authors.forename: Na
authors.initials: N
authors.lastname: Li
authors.affiliation: College of Marine Life Sciences, Ocean University of China, Qingdao, 266003, China. yxmao@ouc.edu.cn.|Key Laboratory of Marine Genetics and Breeding of Ministry of Education, Ocean University of China, Qingdao, 266003, China. yxmao@ouc.edu.cn.
authors.forename: Yunxiang
authors.initials: Y
authors.lastname: Mao
mesh_terms: D021901:DNA, Intergenic; D005822:Genetic Vectors; D030821:Plants, Genetically Modified / Q000235:genetics; D018087:Plastids / Q000235:genetics*; D000461:Rhodophyta / Q000235:genetics*; D012624:Seaweed / Q000235:genetics*; D014170:Transformation, Genetic
chemical_list: D021901:DNA, Intergenic
keywords: Homologous recombination; Plastid transformation; Pyropia yezoensis; psbA UTR