Hi. Does anyone have a cDNA library from Cryptosporidium that I can use?
I am hoping to express Cryptosporidium parvum antigens in the cellular slime
mould Dictyostelium discoideum. We have an established expression system based
on the homologous D. discoideum plasmid Ddp2 and I am presently concentrating on
developing a high transformation frequency suitable for expressing libraries. The
idea is to ligate cDNA into our vectors behind a D. discoideum promoter and
screen for antigen positive cells on a fluorescent activated cell sorter.
D. discoideum is a protozoa and has A+T rich DNA so it may be ideal for
expressing authentic parasite antigens. We can secrete 10-20mg recombinant
protein per litre and it is also easy to grow large quantities of D. discoideum
cells (Dettrich et al, 1994 Bio/technology 12, 614-618). So this expression
system could have lots of applications in parasitology if we can demonstrate the
production and expression of large cDNA libraries.
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