infection - retroviral infection (May/31/2006 )
does anyone can explain me something about retroviral infection? thanks in advance.
the general protocol for retroviral infection normaly performed by 3 round infection to increase the efficiency. my question is after infection, how many copies your interested gene could be intergrated into genome. in my opinion, one copy. according to my knowledge, after infection the host obtained immunity to against being infected by the same virus. this is the case for phage and some virus. but for the geneticaly manipulated retroviral vector, whether it still give the host such kind of immunity, i have no idea.
if there is no host immunity, even we do titration try to keep one cell one virus, by the repeated infection process, we could not gurantee one cell one virus?
i infected cells with WT and 2 point mutation, i always have high expression level with 2 point mutation but not WT. is this due to different copy numbers? or due to compensation of lsot WT expression?
hi
what i know at time, is that infection is done only once per cell. Multiple infections helps obtaining a higher number of infected cells.
what i know at time, is that infection is done only once per cell. Multiple infections helps obtaining a higher number of infected cells.
hi,
I had read from some paper that some reserchers infect cells at low MOI, 0.3 for example, to ensure that each infected cell will receive just one copy of the transgene, while most cells are not infected. Does this mean that at higher MOI, one cell can be infected several times?
thanks for your reply. the protocol of Nolan's lab is 3 times reinfection. this will leading to more copies one cell or increasing infection efficiency
By using a low titres of virus, u would mostly have one virus per cell but there is no guarantee for that. as u might have some cells with more than 1 viral particles for different reasons.
Yes, if u infect with a lot of virus particles, then u could have more than one viral particle per cell. This can be seen when the u infect using a virus coding for fluorescent protien. There are different grades of fluorescence. I assume it is due to more particles per cell.
I have another question. By conventional transfection, there may be hundreds or thousands copies of gfp entering a cell. Is single copy of gfp enough to make cell fluorescent under fluorescent microscope?
hi
single gfp is enough to make a good fluorescence visible in microscopy.
For virus infection : as the integration in the genome is part targeted, it's mostly a unique event for 1cell.
thanks.
according to Fred, if one cell only one copy of virus, how to explain the difference expression level between WT and point mutation. because of compensation?
the phage lysogenic circle and HIV infected human support the idea of Fred: one cell one virus. but we don't know this immunity occurs by the which virus's protein and is this protein coding sequence still kept in the genetic manipulated vector