“…An indication of the re latedness of the clones was that their 3' untranslated re gions, which usually show the highest amount of polymor phism in cDNA [19,20], were identical. The degree of sequence variation described here is not usually found in DNA coding other eukaryotic proteins regions [21] and Kreitman [22] has for example reported that 11 different isolates of Drosophila melanogaster had only one amino acid change in their alcohol dehydrogenase genes.…”
Sequence information has been obtained from 5 cDNA clones encoding the major house dust mite allergen Der p I, including one which codes for the full length preproenzyme form of the molecule. All translated sequences were unique with 1–3 differences between each clone. 4/5 of the substitutions found were to residues found in Der f l and two positions had a substitution in more than one clone. The polymorphisms included residues already described to be in T cell epitopes, so as well as being necessary to consider them in the construction of synthetic allergens, the substitutions will be important for the interpretation of experimental and genetic studies.
“…An indication of the re latedness of the clones was that their 3' untranslated re gions, which usually show the highest amount of polymor phism in cDNA [19,20], were identical. The degree of sequence variation described here is not usually found in DNA coding other eukaryotic proteins regions [21] and Kreitman [22] has for example reported that 11 different isolates of Drosophila melanogaster had only one amino acid change in their alcohol dehydrogenase genes.…”
Sequence information has been obtained from 5 cDNA clones encoding the major house dust mite allergen Der p I, including one which codes for the full length preproenzyme form of the molecule. All translated sequences were unique with 1–3 differences between each clone. 4/5 of the substitutions found were to residues found in Der f l and two positions had a substitution in more than one clone. The polymorphisms included residues already described to be in T cell epitopes, so as well as being necessary to consider them in the construction of synthetic allergens, the substitutions will be important for the interpretation of experimental and genetic studies.
“…X53859; Garber et al, 1990), and a 70% match with the mouse, Mus musculus, CK gene (GenBank Accession No. M74149; van Deursen et al, 1992). Using these alignments we determined the linear arrangement of the belonid exon/intron positions for the approximately 1000 bp CK fragment amplified by the CK1 and CK2 primers.…”
“…A specific antisense oligonucleotide probe complementary to nucleotides 3787-3837 of the coding sequence of murine CKB gene [26] was radioactively labeled, purified, hybridized, and detected as before [27].…”
To screen for new region-specific protein markers we compared the proteome maps of the primary visual and somatosensory areas V1 and S1 in mouse brain using 2-D difference gel electrophoresis (2-D DIGE). Twenty-three protein spots showed a statistically significant difference in expression level between V1 and S1, with 52% appearing more abundantly in V1. Twenty-six proteins were mass spectrometrically identified in 22 spots. To assess the validity of this list of potential areal markers generated by 2-D DIGE, the effective area-specific distribution profile of creatine kinase brain subtype (CKB), a protein with a clearly higher expression level in S1, was monitored with in situ hybridization. The mRNA expression profile of CKB displayed a clear area-specific distribution, which allowed demarcation of S1 and its topographical borders with neighboring neocortical areas. This proteomic study demonstrates the innovative application of 2-D DIGE and MS to select new regional markers for neuroscience research.
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