pTet on advanced载体说明书
- 上海起发实验试剂有限公司2013年7月30日 13:07 点击：2201
Tet-On® Advanced Inducible Gene Expression Systems （pTet on advanced，pTRE Tight，pTRE Tight Luciferase）使用说明书
Vectors： pTet on advanced，pTRE Tight，pTRE Tight Luciferase
下载地址：http://www.chem17.com/st178406/Info_104233.html
A. Summary
The Tet-On Advanced Inducible Gene Expression System is a tightly regulated and highly responsive system that
produces on-demand, robust expression of your gene of interest （GOI） in target cells. The system is established in
target cells by sequentially transfecting them with the provided vectors and selecting stable cell lines. Target cells that
express the Tet-On Advanced transactivator, and that also contain an integrated TRE-based expression vector （e.g.,
pTRE-Tight） will express high levels of your GOI when cultured in the presence of the system’s inducer, doxycycline
（Dox） （Figure 1）.
B. Elements of Tet-On Advanced Induction
Based on the original tetracycline （Tc）-regulated transcriptional transactivators described by Gossen & Bujard
（1992） and Gossen et al. （1995）, Tet-On Advanced is a modified transactivator protein that is optimized for expression in mammalian cells, and which demonstrates higher sensitivity and fidelity than previous versions （Urlinger, et
al. 2000）. The inducible promoter, P
Tight, provides for very low basal expression and tightly controlled induction.
• The Tet-On Advanced transactivator. The pTet-On Advanced vector constitutively expresses the tetracyclinecontrolled transcriptional transactivator, Tet-On Advanced （Urlinger et al., 2000）. This engineered protein
consists of a mutant E. coli TetR protein （rTetR） fused to three minimal "F”-type activation domains derived
from the herpes simplex virus VP16 protein （Baron et al., 1997, Triezenberg et al., 1988）. In the presence
of Dox, Tet-On Advanced binds to the tetO sequences in P
Tight, and activates high level transcription from
this inducible promoter. The Tet-On Advanced coding sequence is fully synthetic and utilizes human codon
preferences to increase its expression level and stability in mammalian cells.
• The P
Tight inducible promoter. This is an inducible promoter that controls transcription of your GOI. The
P
Tight composite promoter was originally developed as the Ptet-14 promoter in the laboratory of Dr. H. Bujard
and consists of a modified Tet-Responsive Element （TREmod） containing 7 direct repeats of the tet operator
sequence, tetO, which is joined to a minimal CMV promoter （P
minCMV?）. P
Tight lacks binding sites for endogenous mammalian transcription factors, so it is virtually silent in the absence of induction. In the presence of
Dox, Tet-On Advanced binds tightly and specifically to P
Tight and activates transcription of the downstream
GOI （Figure 1）.
质粒图谱:
全年征稿 / 资讯合作
联系邮箱：kefu@labbase.net
版权与免责声明
- 凡本网注明“来源：来宝网”的所有作品，版权均属于来宝网，转载请必须注明来宝网， http://www.labbase.net，违反者本网将追究相关法律责任。
- 本网转载并注明自其它来源的作品，目的在于传递更多信息，并不代表本网赞同其观点或证实其内容的真实性，不承担此类作品侵权行为的直接责任及连带责任。其他媒体、网站或个人从本网转载时，必须保留本网注明的作品来源，并自负版权等法律责任。
- 如涉及作品内容、版权等问题，请在作品发表之日起一周内与本网联系，否则视为放弃相关权利。
