title: Successful expression of heterologous egfp gene in the mitochondria of a photosynthetic eukaryote Chlamydomonas reinhardtii.
abstract: The efficient expression of exogenous gene in mitochondria of photosynthetic organism has been an insurmountable problem. In this study, the pBsLPNCG was constructed by inserting the egfp gene into a site between TERMINVREP-Left repeats and the cob gene in a fragment of mitochondrial DNA of Chlamydomonas reinhardtii CC-124 and introduced into the mitochondria of respiratory deficient dum-1 mutation of C. reinhardtii CC-2654. Sequencing and DNA Southern analyses revealed that egfp gene had been integrated into the mitochondrial genome of transgenic algae as expected and no other copy of egfp existed in their nucleic genome. Both the fluorescence detection and Western blot analysis confirmed the presence of eGFP protein in the transgenic algae; it indicated that the egfp gene was successfully expressed in the mitochondria of C. reinhardtii.
authors.affiliation: Shenzhen Key Laboratory of Marine Bioresource and Eco-environmental Science, College of Life Science, Shenzhen University, Shenzhen 518060, PR China. huzl@szu.edu.cn
authors.forename: Zhangli
authors.initials: Z
authors.lastname: Hu
authors.forename: Zhonglin
authors.initials: Z
authors.lastname: Zhao
authors.forename: Zhihua
authors.initials: Z
authors.lastname: Wu
authors.forename: Zhun
authors.initials: Z
authors.lastname: Fan
authors.forename: Jun
authors.initials: J
authors.lastname: Chen
authors.forename: Jinxia
authors.initials: J
authors.lastname: Wu
authors.forename: Jiancheng
authors.initials: J
authors.lastname: Li
mesh_terms: D015139:Blotting, Southern; D015153:Blotting, Western; D016825:Chlamydomonas reinhardtii / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D015870:Gene Expression; D005822:Genetic Vectors; D054629:Genome, Mitochondrial; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics*; D018613:Microscopy, Confocal; D008928:Mitochondria / Q000235:genetics* / Q000378:metabolism; D010788:Photosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics*; D017422:Sequence Analysis, DNA; D014170:Transformation, Genetic
chemical_list: D011994:Recombinant Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins