Enhanced green fluorescent protein (egfp) gene expression in Tetraselmis subcordiformis chloroplast with endogenous regulators
Enhanced green fluorescent protein (egfp) gene expression in Tetraselmis subcordiformis chloroplast with endogenous regulators
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内源调节剂增强 Tetraselmis subcordiformis 叶绿体中绿色荧光蛋白 (egfp) 基因表达
DOI：
10.1007/s11274-016-2039-y
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发表时间：
2016-04
影响因子：
4.1
通讯作者：
Qin Song
中科院分区：
文献类型：
--
作者：
Cui Yulin;Zhao Jialin;Hou Shichang;Qin Song
本研究在基础遗传转化技术的基础上，利用内源调节剂优化亚心形油茶的叶绿体转化。Rrn16S、rbcL、PSBA和PSBC基因在叶绿体中普遍高表达，这些基因的调控常用于叶绿体转化。由于缺乏已知的叶绿体基因组序列，本文采用基因组步移法获得亚心形海棠内源调控基因的全序列。将含有3个启动子、2个终止子和1个核糖体组合序列的启动子插入到以绿色荧光蛋白基因为报告基因的原核表达载体pPSC-R中，制备了5个叶绿体表达载体。用基因枪法成功地将这些载体转化到亚心形锥虫中，并通过显微镜观察EGFP荧光来检测每种载体的效率。结果表明，这些载体的效率均高于携带外源调节子的载体PPSC-G，其中含有Prrn、PSBA-5‘RE和TpsbA的载体效率最高。本研究为亚心形毛藻提供了一套有效的内源调节剂，为今后对该藻的基础研究奠定了基础。
On the basis of fundamental genetic transformation technologies, the goal of this study was to optimize Tetraselmis subcordiformis chloroplast transformation through the use of endogenous regulators. The genes rrn16S, rbcL, psbA, and psbC are commonly highly expressed in chloroplasts, and the regulators of these genes are often used in chloroplast transformation. For lack of a known chloroplast genome sequence, the genome-walking method was used here to obtain full sequences of T. subcordiformis endogenous regulators. The resulting regulators, including three promoters, two terminators, and a ribosome combination sequence, were inserted into the previously constructed plasmid pPSC-R, with the egfp gene included as a reporter gene, and five chloroplast expression vectors prepared. These vectors were successfully transformed into T. subcordiformis by particle bombardment and the efficiency of each vector tested by assessing EGFP fluorescence via microscopy. The results showed that these vectors exhibited higher efficiency than the former vector pPSC-G carrying exogenous regulators, and the vector pRFA with Prrn, psbA-5'RE, and TpsbA showed the highest efficiency. This research provides a set of effective endogenous regulators for T. subcordiformis and will facilitate future fundamental studies of this alga.
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DOI：
--
发表时间：
1983
期刊：
Developments in biological standardization
影响因子：
--
作者：
N. Harford;T. Cabezón;M. Crabeel;E. Simoen;A. Rutgers;M. de Wilde
通讯作者：
N. Harford;T. Cabezón;M. Crabeel;E. Simoen;A. Rutgers;M. de Wilde
影响因子：
5.2
作者：
Fukuda, Satoru;Mikami, Koji;Saga, Naotsune
通讯作者：
Saga, Naotsune
影响因子：
46.9
作者：
De Cosa, B;Moar, W;Daniell, H
通讯作者：
Daniell, H
影响因子：
2.7
作者：
Guo, Zhen;Chen, Zhaoan;Jin, Meifang
通讯作者：
Jin, Meifang
DOI：
--
发表时间：
1999
期刊：
The Plant journal : for cell and molecular biology
影响因子：
--
作者：
Sidorov;Kasten;Pang;Hajdukiewicz;Staub;Nehra
通讯作者：
Sidorov;Kasten;Pang;Hajdukiewicz;Staub;Nehra