How to overexpress a gene in C.elegans - (Dec/13/2007 )
Hi.I want to study the function of a gene.but I have troubles in overexpressing the gene.
How to overexpress a gene in C.elegans? What plasmids should I use? Who can give me suggests or a prptocol?
-fds-
QUOTE (fds @ Dec 13 2007, 02:15 AM)
Hi.I want to study the function of a gene.but I have troubles in overexpressing the gene.
How to overexpress a gene in C.elegans? What plasmids should I use? Who can give me suggests or a prptocol?
How to overexpress a gene in C.elegans? What plasmids should I use? Who can give me suggests or a prptocol?
Several years ago Andy Fire's lab made a whole series of useful vectors for expression in C. elegans. You can get information about it here:
http://www.addgene.org/pgvec1?f=c&cmd=...col&colid=1
or google Andy Fire and you will find the information. Most C. elegans labs already have these vectors, so once you find what you want I would just ask a C. elegans lab in your vicinity if you can borrow.
-smu2-
hi
i have tried pPD49.83 which is for tissue over expression, and pPD39.80, which is heat shock promote. In my experience, the tissue over expression yield high amount of protein, however very hard to purify. The yield from the heat shock one pretty low, however, the protein seems more soluble.
-GRC_YEO-
QUOTE (GRC_YEO @ Dec 13 2007, 07:25 PM)
hi
i have tried pPD49.83 which is for tissue over expression, and pPD39.80, which is heat shock promote. In my experience, the tissue over expression yield high amount of protein, however very hard to purify. The yield from the heat shock one pretty low, however, the protein seems more soluble.
i have tried pPD49.83 which is for tissue over expression, and pPD39.80, which is heat shock promote. In my experience, the tissue over expression yield high amount of protein, however very hard to purify. The yield from the heat shock one pretty low, however, the protein seems more soluble.
Hi
i do not know which plasmid can express in all tissures. pPD49.83 seems to express in muscle and hypodermis . Any plasmid can express in as many as tissures?
-fds-