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Transgenic mouse milk

b-choi2 at STUDENTS.UIUC.EDU b-choi2 at STUDENTS.UIUC.EDU
Sat Sep 30 21:59:19 EST 1995


Hello,

I am working on bovine beta casein which is one of milk proteins.  This
bovine beta casein (wild-type) does not have any potential N-linked
glycosylation sites in its stuctural gene.  For some reason we introduced
N-glycosylation site (Asn-X-Ser) into bovine beta casein gene and generated
transgenic mice carrying bovine beta casein gene (wild-type or mutant
containing single glycosylation site).  We found glycosylated and
unglycosylated bovine beta casein in the milk of transgenic mice using
bovine beta casein specific antibody and confirmed N-linked glycosylation
using PNGase F treatment.  However, I have difficulty in detecting
glycosylated beta casein using lectins.   The lectin blots with Con A, WGA,
ECA and UEA-1 did not show any positive reactions.  So I am wondering what
the reason is.  If N-linked oligosaccharides are sulfated, what kind of
method can I try to detect N-linked sugar ?

Thank you.

Byung-Kwon Choi  






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