caa a22 4500 397573227 CHVBK 20180308164847.0 cr unu---uuuuu 161202e199606 xx s 000 0 eng 10.1093/hmg/5.6.771 doi (NATIONALLICENCE)oxford-10.1093/hmg/5.6.771 Cloning and Expression of the Gene Involved in Sanfilippo B Syndrome (Mucopolysaccharidosis III B) [Elektronische Daten] [Birgit Weber, Lianne Blanch, Peter R. Clements, Hamish S. Scott, John J. Hopwood] Sanfilippo B syndrome is caused by a deficiency of α-N-acetylglucosaminidase, a lysosomal enzyme involved in the degradation of heparan sulphate. Accumulation of the substrate in lysosomes results in degeneration of the central nervous system with progressive dementia often combined with hyperactivity and aggressive behaviour. In order to clone the deficient gene, we purified the enzyme from human placenta and obtained amino acid sequence information. Alignment of one of the CNBr generated internal peptides to sequence from the database revealed the chromosomal location of the gene in the 5′ upstream flanking region of the gene for 17-β-hydroxysteroid-dehydrogenase at 17q21.1. The available DNA sequence was used to clone the cDNA coding for α-N-acetylglucosaminidase and analyse its gene structure. The gene is fully contained in the 5′ upstream flanking region of the gene for 17-β-hydroxysteroid-dehydrogenase and interrupted by five introns. The cDNA clone has a length of 2575 bp and encodes a protein of 743 amino acids. Chinese hamster ovary cells transfected with the cDNA construct show α-N-acetylglucosaminidase activity about 17-fold over background. This will allow correction studies with NAG deficient Sanfilippo B cell lines and facilitate the development of enzyme replacement therapy for these patients. © 1996 Oxford University Press Weber Birgit Lysosomal Diseases Research Unit, Department of Chemical Pathology, Women's and Children's Hospital, North Adelaide S.A. 5006, Australia aut Blanch Lianne Lysosomal Diseases Research Unit, Department of Chemical Pathology, Women's and Children's Hospital, North Adelaide S.A. 5006, Australia aut Clements Peter R. Lysosomal Diseases Research Unit, Department of Chemical Pathology, Women's and Children's Hospital, North Adelaide S.A. 5006, Australia aut Scott Hamish S. Lysosomal Diseases Research Unit, Department of Chemical Pathology, Women's and Children's Hospital, North Adelaide S.A. 5006, Australia aut Hopwood John J. Lysosomal Diseases Research Unit, Department of Chemical Pathology, Women's and Children's Hospital, North Adelaide S.A. 5006, Australia aut Human Molecular Genetics Oxford University Press 5/6(1996-06), 771-777 0964-6906 5:6<771 1996 5 hmg https://doi.org/10.1093/hmg/5.6.771 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/hmg/5.6.771 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Weber Birgit Lysosomal Diseases Research Unit, Department of Chemical Pathology, Women's and Children's Hospital, North Adelaide S.A. 5006, Australia aut NATIONALLICENCE 700 1- Blanch Lianne Lysosomal Diseases Research Unit, Department of Chemical Pathology, Women's and Children's Hospital, North Adelaide S.A. 5006, Australia aut NATIONALLICENCE 700 1- Clements Peter R. Lysosomal Diseases Research Unit, Department of Chemical Pathology, Women's and Children's Hospital, North Adelaide S.A. 5006, Australia aut NATIONALLICENCE 700 1- Scott Hamish S. Lysosomal Diseases Research Unit, Department of Chemical Pathology, Women's and Children's Hospital, North Adelaide S.A. 5006, Australia aut NATIONALLICENCE 700 1- Hopwood John J. Lysosomal Diseases Research Unit, Department of Chemical Pathology, Women's and Children's Hospital, North Adelaide S.A. 5006, Australia aut NATIONALLICENCE 773 0- Human Molecular Genetics Oxford University Press 5/6(1996-06), 771-777 0964-6906 5:6<771 1996 5 hmg Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford