title: Construction of rice site-specific chloroplast transformation vector and transient expression of EGFP gene in Dunaliella salina.
abstract: Chloroplast is a new hotspot in the field of plant transformation system of plant genetic engineering. Plastid transformation has several advantages: high expression, multiple expressed genes in a single transformation event, absence of gene silencing, et al. A series of elements for construction of dicistronic site-specific integration expression vector of rice chloroplast have been cloned, including trnl-trnA (rice chloroplast homologous recombination fragments), Prrn (16S rRNA operon promotor), PpsbA (the 3' untranslated region of the chloroplastpsbA gene), hptll gene (encoding hygromycin phosphotransferase) and EGFP (encoding enhanced green fluorescence protein). All the elements were constructed into a rice chloroplast dicistronic expression vector pCTE04 (-trnl-Prrn-RBS-hptlI-RBS-EGFP-PpsbA- trnA-). Then pCTE04 was introduced into chloroplasts of Dunaliella salina through particle bombardment. Strong green fluorescence was observed in chloroplasts of some bombarded Dunaliella salina cells under a stereo fluorescence microscope, indicating that pCTE04 could be expressed in Dunaliella salina chloroplasts transiently. It provides a solid foundation for further genetic engineering in rice chloroplast transformation.
authors.affiliation: Longping Branch, Graduate School of Central South University, Changsha 410125, China.
authors.forename: Ding
authors.initials: D
authors.lastname: Li
authors.forename: Xiaoxia
authors.initials: X
authors.lastname: Han
authors.forename: Jia
authors.initials: J
authors.lastname: Zuo
authors.forename: Lingling
authors.initials: L
authors.lastname: Xie
authors.forename: Ronghua
authors.initials: R
authors.lastname: He
authors.forename: Jing
authors.initials: J
authors.lastname: Gao
authors.forename: Lan
authors.initials: L
authors.lastname: Chang
authors.forename: Longping
authors.initials: L
authors.lastname: Yuan
authors.forename: Mengliang
authors.initials: M
authors.lastname: Cao
mesh_terms: D002736:Chloroplasts / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular / Q000379:methods; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D012275:Oryza / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000235:genetics; D016823:Volvocida / Q000235:genetics* / Q000378:metabolism
chemical_list: C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins