Expression of the tandem enhanced yellow fluorescent marker gene in Toxoplasma gondii
Expression of the tandem enhanced yellow fluorescent marker gene in Toxoplasma gondii
复制标题
串联增强黄色荧光标记基因在弓形虫中的表达
DOI：
10.1007/s00436-009-1448-3
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发表时间：
2009-05
影响因子：
2
通讯作者：
Li, Xuelian
中科院分区：
文献类型：
--
作者：
Zhang, Wei;Chen, Dan;Hao, Yongxin;Liu, Qun;Ding, Jun;Li, Xuelian
荧光蛋白是监测基因表达和研究包括寄生虫在内的生物体生物学过程的有用工具。为了提高弓形虫的转染效率和荧光强度，构建了一种新的弓形虫致密颗粒蛋白1（GRA 1）启动子控制的串联增强型黄色荧光蛋白（EYFP-EYFP）瞬时表达载体RGES。将RGES质粒导入T.电穿孔法制备弓形虫RH株。用EYFP-EYFP基因转染高比例的速殖子（约25.5%），并且在电穿孔后48-72 h检测到高荧光强度。通过聚合酶链反应（PCR）、逆转录酶PCR和Western blotting证实串联EYFP基因及其在转染子中的表达。EYFP-EYFP的转染效率和荧光强度均高于单一EYFP或绿色荧光蛋白。EYFP-EYFP基因是研究T.刚地。
Fluorescent protein is a useful tool for monitoring gene expression and studying biological processes of organisms including parasites. To improve the transfection efficiency and fluorescent intensity in Toxoplasma gondii, a new transient expression vector, RGES, containing the tandem enhanced yellow fluorescent protein gene, EYFP-EYFP, under the control of the parasite dense granule protein 1 (GRA1) promoter was constructed. The RGES plasmid was introduced into T. gondii RH strain by electroporation. A high proportion of tachyzoites (approximately 25.5%) was transfected with the EYFP-EYFP gene and high fluorescence intensity was detected 48–72 h after electroporation. The tandem EYFP gene and its expression in the transfectants were confirmed by polymerase chain reaction (PCR), reverse transcriptase PCR, and Western blotting. The efficiency of transfection and fluorescent intensity of EYFP-EYFP were greater than that of the single EYFP or green fluorescent protein. The EYFP-EYFP gene is a better marker for the study of biological processes of T. gondii.
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影响因子：
1.5
作者：
B. Striepen;C. He;M. Matrajt;D. Soldati;D. Roos
通讯作者：
B. Striepen;C. He;M. Matrajt;D. Soldati;D. Roos
影响因子：
4
作者：
Nishikawa, Y;Xuan, XN;Nagasawa, H
通讯作者：
Nagasawa, H
影响因子：
56.9
作者：
CHALFIE, M;TU, Y;PRASHER, DC
通讯作者：
PRASHER, DC
DOI：
10.1073/pnas.91.12.5508
发表时间：
1994-06
影响因子：
11.1
作者：
L. Sibley;M. Messina;I. Niesman
通讯作者：
L. Sibley;M. Messina;I. Niesman
影响因子：
13.8
作者：
CUBITT, AB;HEIM, R;TSIEN, RY
通讯作者：
TSIEN, RY