title: Chloroplast transformation of Platymonas (Tetraselmis) subcordiformis with the bar gene as selectable marker.
abstract: The objective of this research was to establish a chloroplast transformation technique for Platymonas (Tetraselmis) subcordiformis. Employing the gfp gene as a reporter and the bar gene as a selectable marker, transformation vectors of P. subcordiformis chloroplast were constructed with endogenous fragments rrn16S-trnI (left) and trnA-rrn23S (right) as a recombination site of the chloroplast genome. The plasmids were transferred into P. subcordiformis via particle bombardment. Confocal laser scanning microscopy indicated that the green fluorescence protein was localized in the chloroplast of P. subcordiformis, confirming the activity of the Chlamydomonas reinhardtii promoter. Cells transformed with the bar gene were selected using the herbicide Basta. Resistant colonies were analyzed by PCR and Southern blotting, and the results indicated that the bar gene was successfully integrated into the chloroplast genome via homologous recombination. The technique will improve genetic engineering of this alga.
authors.affiliation: Key Laboratory of Coastal Biology and Biological Resource Utilization, Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Yantai, Shandong, China; Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao, 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, Yantai, Shandong, China.
authors.forename: Song
authors.initials: S
authors.lastname: Qin
authors.affiliation: Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Jiang
mesh_terms: D000123:Acetyltransferases / Q000235:genetics*; D000460:Chlorophyta / Q000235:genetics*; D002736:Chloroplasts / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods*; D005819:Genetic Markers / Q000235:genetics; D005974:Glutamate-Ammonia Ligase / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D014170:Transformation, Genetic
chemical_list: D005819:Genetic Markers; D000123:Acetyltransferases; C056243:phosphinothricin N-acetyltransferase; D005974:Glutamate-Ammonia Ligase