I found there wasn't termination for antibiotic resistant gene transcription in many vectors.
I think stopless transcription will affect activation of following replication origin or another promoter. Is it true? or false? or not known?
Does anyone know about the answer?
Furthermore, if anyone know the sequence of Ampicillin termination site, or substitute sequence, please let me know.
Yes, runaway transcription will interfere with (but not eliminate) plasmid replication and/or transcription of genes in opposite orientation. However, how have you determined that these genes have no transcriptional terminator? Is this experimental data, or have you just scanned the sequence for 3' stem and loops followed by T-rich streteches?
Thanks Martin for your answer.
Absence of termination is not confirmed by experiment. I just read information sheets of some vectors, and I confirmed any sheet doesn't say about termiation for resistant gene. Thus, it is possible that vectors have termination, even though it is not mentioned in information sheet.
Furthermore, i investigated downstream sequences of resistant gene from diverse type of vectors. They differ each other, even though promoter sequence of resistant gene from those vectors are same. Thus, i concluded they have promoter for resistant gene, but don't have termination for resistant gene. (it might be false.)
Do you know reliable software program or method to identify termination sequence? (such as program for identifying T-rich streteches.)
There's two I know of; one is a web server called ARNold at:
http://rna.igmors.u-psud.fr/toolbox/arnold/
And there is TransTerm:
http://transterm.cbcb.umd.edu/
But the problem is that they are only good for rho-independent terminators, so there is always a chance that the markers are using rho-dependent termination (which depend on rut sites within the coding sequence).
Thanks for your help!
ARNold is very useful. It show some vectors have rho-independent terminator for resistant gene and some vectors not. I solved my question! I appreciate for your help.
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