Based on the partial amino acid sequence, cDNA encoding UDP-GalNAc:Fuc alpha 1----2Gal 1----3GalNAc transferase, specific primary gene product of histo-blood group A (A transferase), was cloned and sequenced. Poly(A)+ RNA from human stomach cancer cell line MKN45, expressing high levels antigen, used for construction a lambda gt10 library. Degenerate synthetic oligodeoxynucleotides were polymerase chain reactions to detect presence sequence interest in (presence test) identify correct clones...

A significant percentage of human tumors contain activated ras oncogenes that have acquired oncogenic potential as a result somatic point mutations at codon 12 or 61 the encoded gene product. We report here method to detect and characterize in genes is based on ability pancreatic ribonuclease (RNase A; EC 3.1.27.5) cleave RNA heteroduplexes containing single-base mismatches. Using this method, we show certain tumor cells mutant c-Ki-ras genes, define nature position these mutations. At same...

Four amino acid substitutions (aa 176, 235, 266, and 268) have been found between the coding regions of cDNAs for histo-blood group A B transferases (Yamamoto F., Clausen, H., White, T., Marken, J., Hakomori, S. (1990) Nature 345, 229-233). Here we establish basis differential affinity these glycosyltransferases to nucleotide-sugar (UDP-GalNAc or UDP-Gal). On gene reconstruction experiments studies expression in DNA transfected HeLa cells, third as well fourth aa (leucine glycine methionine...

We have isolated human genomic DNA clones encompassing 30 kbp of the histo-blood group ABO locus. The locations exons been mapped and nucleotide sequences exon-intron boundaries determined. genes consist at least seven exons, coding sequence in spans over 18 kb DNA. range size from 28 to 688 bp, with most lying exon 7.

There are three common alleles (A, B, and O) at the human ABO blood group locus. We compared nucleotide sequences of these alleles, relatively large numbers differences were found among them. These correspond to divergence time least a few million years, which is unusually for allelic under neutral evolution. constructed phylogenetic networks nonhuman primate independent appearances B from ancestral A form observed. results suggest that some kind balancing selection may have been operating...

Abstract We have determined the nucleotide sequence of coding region in last two exons ABO genes from cis ‐AB individuals (genotype ‐AB/O) with no consanguinity. In this region, alleles these 2 were identical to one another while different A 1 allele by substitutions. Both substitutions result amino acid The first substitution is previously found allele. other at fourth position four which discriminate and B transferases.

Human lung tumors PR310 and PR371 maintained in nude mice contain activated c-K-ras oncogenes detectable by the ability of their DNAs to induce morphological transformation NIH 3T3 mouse fibroblasts. Using phage libraries constructed with DNA from fibroblast transformants, we have isolated human sequences that span greater than 40 kilobase pairs oncogene. Based on conservation these conclude transforming oncogene resides within a 43- 46-kilobase-pair region. No clear differences were...

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This year commemorates the 100 th anniversary of discovery ABO blood group system by Karl Landsteiner. His findings red cell agglutination serum and recognition groups laid scientific basis for safe practice transfusion. Even though dozens systems have been identified, still remains to be one most important in transfusion medicine. In 1990, we elucidated molecular genetic three major alleles at locus. Since then witnessed progress our understanding genes A B glycosyltransferases specified a...

Abstract We have encountered an allele which seems to be another type of O at the human histo‐blood group ABO locus. determined nucleotide sequence this over coding region in last two exons. This does not possess single‐nucleotide deletion found common among all alleles previously analyzed. Compared with A 1 allele, has three substitutions resulting amino acid substitutions. The introduction these into transferase expression construct apparently abolished enzymatic activity transferase.

Abstract We have determined the nucleotide sequences of coding region in last two exom ABO genes (which occupy 91% soluble form A 1 transferase) from 7 individuals with weak subgroup phenotypes. Four had an 3 phenotype and a B phenotype. based on PCR followed by subcloning DNA sequencing amplified fragments. Two cases allele case were found to contain single‐base substitution which resulted amino acid substitution. However, no other alleles differences this region. This finding demonstrates...

Abstract von Willebrand factor (vWF) is a multimeric plasma protein with ABO (H) blood group sugar chains. We investigated total of 330 plasmas from normal individuals having various genotypes, special reference to vWF antigen and its platelet glycoprotein‐Ib‐related biological activities, termed ristocetin cofactor (RCof) botrocetin (BCof). RCof reflects the activity higher multimers, while BCof that multimers all sizes. Plasmas carrying one O gene (genotypes AO BO) had slightly, but...

Abstract We have employed a PCR approach to determine the nucleotide sequences of coding region in last two exons histo‐blood group ABO genes from one A x and B(A) individual. Compared with 1 alleles, allele has single substitution (T→ at 646) resulting an amino acid (phenylalanine→isoleucine 216). B (A) substitutions (T→C nt. 657 A→G 703) (serine→glycine aa. 235). The this is located second four which discriminate human transferases, residue (glycine) identical that transferase suggesting...

The ABO system is one of the most important blood group systems in transfusion/transplantation medicine. However, evolutionary significance gene and its polymorphism remained unknown. We took an integrative approach to gain insights into process genes, including those related not only phylogenetically but also functionally. experimentally created a code table correlating amino acid sequence motifs gene-encoded glycosyltransferases with GalNAc (A)/galactose (B) specificity assigned A/B...

Abstract Background ABO blood grouping compatibility restrictions present one of the first hurdles to recipient-donor matching in transplantation. While group O individuals would usually be described as ‘universal’ donors, a rare called Bombay type is incompatible even with O. Here, we describe use bacterial fucosidase enzyme FucOB convert human kidneys during ex vivo hypothermic machine perfusion (HMP). Methods Four declined for transplantation and offered research were recruited this...

Abstract Introduction Enzymatic blood group antigen conversion is a promising strategy to enable ABO-incompatible kidney transplantation. An enzyme called FucOB can cleave the H-antigen, precursor all structures. When applied during ex vivo machine perfusion it could provide an ABO compatible option for rare Bombay recipients. However, this provides only phenotype alteration of antigens on vascular endothelial cells. As these cells remain genetically unmodified and metabolically active,...

We have investigated the regulatory role of DNA methylation in expression human histo-blood group ABO genes. The gene promoter region contains a CpG island whose status correlates well with cell lines tested. was found hypomethylated some that expressed genes, whereas other did not express genes were hypermethylated. Whereas constitutive transcriptional activity demonstrated both expressor and nonexpressor by transient transfection reporter constructs containing sequence, <i>Hha</i>I...

Histo-blood group A transferase produces antigens and transfers GalNAc to the acceptor substrate, H structures of glycolipids glycoproteins. B galactose in place same substrate synthesize antigens. We have previously identified four amino acid substitutions between human transferases. Out these four, at last two positions (codons 266 268) were found be crucial for different donor nucleotide-sugar specificities transferases as analyzed by gene transfer chimeric A-B genes.In present study, we...