kDNA minicircles - forwarded

[Douglas Barker]

>>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
>>
IN THE OLDER LITERATURE I REMEMBER A PAPER 1970'S BY KEN WOLSTENHOLE IN
UTAH WHO HAD AN OPEN READING FRAME IN CRITHIDIA , DAVID ARNOT NOW AT - CELL
AND IMMUNOBIOLOGY - EDINBURGH UNIVERSITY  ALSO FOUND AN OPEN READING FRAME
IN LEISHMANIA - PAHO WORKSHOP 1980 - PUBLISHED AS BIOCHEMICAL
CHARACTERISATION OF LEISHMANIA IN 1982 - AND I AM PRETTY SURE THAT JOSEPH
SHOMLAI AT THE KUVIN CENTRE IN JERUSALEM  ALSO HAS PUBLISHED FAIRLY
RECENTLY ON PROTEIN CODING OF KDNA. SORRY I DO NOT HAVE TIME TO LOOK THEM
ALL UP BUT EMAIL TO THESE PEOPLE MAY HELP YOU. OUR OWN DATA BASE ON THE WWW
CONTAINS ALL THE MINICIRCLE SEQUENCES WE CAN DREDGE OUT OF THE
LITERATURE.(www.ebi.ac.uk/parasites/kDNA/Source.html) CONTACT SUSAN
BREWSTER AT  sb10005 at mole.bio.cam.ac.uk.
YOURS
DOUGLAS C. BARKER




YOU WROTE
>>        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)
>>