length of reporter gene - (May/15/2006 )
do i need to use full length reporter gene to make an expresseion cassette??
wht happen if I use 1200 bp of reporter gene whether its full length is 1800 bp???
I think that it depends on if the gene is part of a gene family.
If it is, then you’ll need a long one in order to be able to distinguish between different members of the family.
If not then in my mined there is no problem in having it even shorter.
sorry i cant understand wht do u mean by "if the gene is part of a gene family"
my purpose is in the attached file and my question is wht happen if I make shorter the "b" gene
i cant manage the enzyme cut sites to make the expression cassette, so i want to shorter the b gene so that i can get suitable enzyme cut option
sorry i send the previous mail without attaching the file. here i attached the file where u can understand my purpose
Has anyone detected the reporter when u having used 1200bps of it instead of 1800bps. If others could detect then its fine else you might need to clone the whole thing in.
If you dont have an option with restriction sites , you could amplifying it with specific sites.
according to my previous attachment the length of b gene is 2600 bp
i want to make NcoI (CCATGG) site in ATG of b gene
but already there is 2 NcoI site is exists in 185nt and 537 nt. so I want to make short the b gene cutting by NcoI to get the 537 -2600 bp fragment. but i donno whether it is possible for my expt or not
Is there a digestion site that is compatible to ligate with the Nco of your cassette or vector? You can also amplify the gene with primers that introduce whatever sites you want to the end of the cassette or integration. So if you have a site other than Nco unique to your ligation that is not in the reporter, you can amplify, cut and ligate with that. Did I understand you situation correctly..?