Hi all,
I am new in lentivirus transduction work and have some problem as listed below:
1) Is CMV promoter considered a strong promoter to express the gene I want in the transgenic organism (said the mammalian cell line which has been inserted with the lentivirus)?
2) For plasmid work, it is known that we can use bacteria transformation to propagate the plasmid. As for virus, is there any way we can propagate the virus in vitro?
Hi Chong,
1) CMV is a good stable mammalian promotor.
2) For virus production this is not possible, the virus is designed in a way it cant recreate new viruses, that of course would be dangerous.
I advise to simply use bigger dishes when transfecting your virus producing cells.
More cells = more medium = higher virus yields.
Jan
1) The CMV promoter is incredibly strong. You will get very high levels of transcription in almost all mammalian cells.
2) Viruses are usually made for research purposes through in vitro culture. You need to transform your host cells with 2-3 plasmids, allow for virus production, and then harvest the virus produced in the media. Depending on the titer desired and target cells that will be transduced, this can be very simple process or it may require an arduous weeks-long process.
- Badges
- Science topic