kDNA minicircles - forwarded

[Janice Taverne]

>From: root at cscdri.ren.nic.in
>>From: Superuser <root at cscdri.ren.nic.in>
>To: j.taverne at ucl.ac.uk
>Date: Tue, 24 Mar 98 8:51:15 PST
>
>        Since I cannot access the parasitology biotech forum
>directly from my end, that's why I am writing to you to
>kindly float my following query for an answer, to parasitology
>colleagues.
>        During the course of my DNA sequencing of kDNA minicircles
>of an isolate of Leishmania, I have come across an open reading
>frame. It is of 93 a.a. and codes for an extremely hydrophobic
>putative protein of 10 kD. An ORF in the variable region of
>minicircles is highly surprising to me as I thought that they do
>not code for proteins. But by northern blot analysis I could even
>detect a transcript! This then prompted me to express the ORF in
>E. coli. I got the expression library with the insert in the right
>orientation etc but I cannot see the protein on SDS-PAGE. Could
>it be that the protein is not finally translated or is its extreme
>hydrophobicity giving a problem? Looking forward eagerly to some
>suggestions.
>        Neeloo Singh (root at cscdri.ren.nic.in)
>