DOI Seite / Zitierlink:
https://doi.org/10.11588/diglit.48120#0073
Abstracts
55
antibodies to a novel HOkD nuclear/cytoplasmic protein with systemic autoim-
mune disease and certain viral infections.
cDNA Cloning and Expression of UlsnRNP C Protein
Y. Misaki1, K. Yamamoto1, H. Miura1, H. Fujii1, K. Nishioka2,
and T. Miyamoto ’
1 Department of Medicine and Physical Therapy, Faculty of Medicine, University of
Tokyo, Japan
2 Institute of Rheumatology, Tokyo Women‘s Medical College, Tokyo, Japan
As previously reported, we have cloned a cDNA termed PS 2 which encodes
UlsnRNP C protein (J. Immunol. 1988). Here we report the cloning and expres-
sion of a putative full lenght cDNA encoding the entire coding region of the
UlsnRNP C protein.
Using PS 2 as a probe, we have screened cDNA libraries constructed from
human fibroblasts by the Okayama-Berg method, which were found to contain
many full-length cDNAs. After screening two different cDNA libraries, the lon-
gest cDNA insert hybridized with PS 2 was 800 bp. We call this cDNA as FS2. In
the Northern blotting using RNA from human PBC, nearly the same length of
the band was identified. Sequencing of FS2 revealed that it contained the initia-
tion codon in the long open reading frame. These results suggest that FS2 is a
full-length cDNA encoding the UlsnRNP C protein, although FS2 could encode
only 15 kDa protein.
In order to express this cDNA in E. coll, we subcloned the insert into an ex-
pression vector, pEX. This vector produces a fusion protein with cro-/?-galac-
tosidase. Transformed E. coli was screened with anti-RNP positive sera. A single
colony, FS2EX was identified to produce the fusion protein which was positive
with the patients’ sera. We are now trying to discriminate the epitope recognized
by anti-RNP antibodies by generating the deletion mutants of this FS2EX.
55
antibodies to a novel HOkD nuclear/cytoplasmic protein with systemic autoim-
mune disease and certain viral infections.
cDNA Cloning and Expression of UlsnRNP C Protein
Y. Misaki1, K. Yamamoto1, H. Miura1, H. Fujii1, K. Nishioka2,
and T. Miyamoto ’
1 Department of Medicine and Physical Therapy, Faculty of Medicine, University of
Tokyo, Japan
2 Institute of Rheumatology, Tokyo Women‘s Medical College, Tokyo, Japan
As previously reported, we have cloned a cDNA termed PS 2 which encodes
UlsnRNP C protein (J. Immunol. 1988). Here we report the cloning and expres-
sion of a putative full lenght cDNA encoding the entire coding region of the
UlsnRNP C protein.
Using PS 2 as a probe, we have screened cDNA libraries constructed from
human fibroblasts by the Okayama-Berg method, which were found to contain
many full-length cDNAs. After screening two different cDNA libraries, the lon-
gest cDNA insert hybridized with PS 2 was 800 bp. We call this cDNA as FS2. In
the Northern blotting using RNA from human PBC, nearly the same length of
the band was identified. Sequencing of FS2 revealed that it contained the initia-
tion codon in the long open reading frame. These results suggest that FS2 is a
full-length cDNA encoding the UlsnRNP C protein, although FS2 could encode
only 15 kDa protein.
In order to express this cDNA in E. coll, we subcloned the insert into an ex-
pression vector, pEX. This vector produces a fusion protein with cro-/?-galac-
tosidase. Transformed E. coli was screened with anti-RNP positive sera. A single
colony, FS2EX was identified to produce the fusion protein which was positive
with the patients’ sera. We are now trying to discriminate the epitope recognized
by anti-RNP antibodies by generating the deletion mutants of this FS2EX.