Construction of Rice Site-Specific Chloroplast Transformation Vector and Transient Expression of EGFP Gene in Dunaliella Salina
文献类型： 外文期刊
作者： Li, Ding 1 ; Han, Xiaoxia 1 ; Zuo, Jia 1 ; Xie, Lingling 1 ; He, Ronghua 1 ; Gao, Jing 1 ; Chang, Lan 3 ; Yuan, Longping; 1 ;
作者机构： 1.Cent S Univ, Grad Sch, Longping Branch, Changsha 410125, Hunan, Peoples R China
2.Hunan Hybrid Rice Res Ctr, State Key Lab Hybrid Rice, Changsha 410125, Hunan, Peoples R China
3.Hunan Longping High Tech Gene Ltd Co, Changsha 410125, Hunan, Peoples R China
关键词： Chloroplast;Dunaliella Salina;EGFP;Transient Expression
期刊名称：JOURNAL OF BIOMEDICAL NANOTECHNOLOGY （ 影响因子：4.099； 五年影响因子：3.492 ）
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收录情况： SCI
摘要： 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-hptll-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.
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