A journey to the world of glycobiology

Finding of the deletion phenomenon of certain oligosaccharides in human milk and its correlation to the blood types of the donors opened a way to elucidate the biochemical basis of blood types in man. This success led to the idea of establishing reliable techniques to elucidate the structures and functions of the N-linked sugar chains of glycoproteins. N-Linked sugar chains were first released quantitatively as oligosaccharides by enzymatic and chemical means, and labelled by reduction with NaB³H₄. After fractionation, structures of the radioactive oligosaccharides were determined by a series of methods developed for the studies of milk oligosaccharides. By using such techniques, structural rules hidden in the N-linked sugar chains, and organ- and species-specific N-glycosylation of glycoproteins, which afforded a firm basis to the development of glycobiology, were elucidated. Finding of galactose deficiency in the N-linked sugar chains of serum lgG from patients with rheumatoid arthritis, and malignant alteration of N-glycosylation in various tumors opened a new research world called glycopathology.However, recent studies revealed that several structural exceptions occur in the sugar chains of particular glycoproteins. Finding of the occurrence of the Gal1-4Fuc1- group linked at the C-6 position of the proximal N-acetylglucosamine residue of the hybrid type sugar chains of octopus rhodopsin is one of such examples. This finding indicated that the fucosyl residue of the fucosylated trimannosyl core should no more be considered as a stop signal as has long been believed. Furthermore, recent studies on dystroglycan revealed that the sugar chains, which do not fall into the current classification of N- and O-linked sugar chains, are essential for the expression of the functional role of this glycoprotein.It was found that expression of many glycoproteins is altered by aging. Among the alterations of the glycoprotein patterns found in the brain nervous system, the most prominent evidence was found in P₀. This protein is produced in non-glycosylated form in the spinal cord of young mammals. However, it starts to be N-glycosylated in the spinal cord of aged animals.These evidences indicate that various unusual sugar chains occur as minor components in mammals, and play important roles in particular tissues.  

单克隆抗体血型定型试剂转瓶培养生产工艺

研制成功单克隆抗体ABO血型定型试剂转瓶培养生产工艺，生产周期从静置培养生产的5～6天缩短到2～3天。产品所有质量指标均达到或超过国家标准，特别是抗体效价明显提高。实验及生产结果显示，该法简单易行，经济快速．投入低，产出高，生产时间短，产品质量高。  

红鲫血型的血清学研究

运用经典血清学方法首次在红鯽中证明了一种与金枪鱼中的A-B-O血型及人类的ABO血型模式相似的红细胞抗原系统,命名为S血型系统。该系统有四种血型表型:S~1,S~2,S~2S~2和S~0,推测它由S~1、S~2和S~0三个复等位基因决定。  

Genetic relationships among European cattle breeds

Genetic relationships among 37 European cattle breeds were investigated using blood group and serum protein polymorphisms. The 18 859 animals included in the study represented a random sample from pedigree populations in the UK. Within-breed variation was estimated by average heterozygosity and number of alleles observed, and breed relationships were evaluated by genetic distance. Standard errors of the heterozygosity, number of alleles and genetic distance were obtained by bootstrapping. The significance of breed differences was tested using an exact test of differentiation. French, Italian and Channel Island breeds were found to have generally higher heterozygosities and a greater number of alleles than breeds from mainland Britain and North Europe. Genetic distances ranged between 0·011 (±0·005) and 0·309 (±0·071). Two major breed groups were identified; a group of French, Italian and Channel Island breeds together with the Simmental and Gelbvieh, and a second group consisting of the mainland British and North European breeds. The exact test of breed differentiation showed all breeds to be significantly different from one another ( P < 0·0001). Overall relationships among breeds reflected their geographical origin and common ancestry rather than the agricultural use for which the breeds have been selected.  

基于氨基酸特征序列对人类Rh血型系统的蛋白质结构分析

利用代数学中同态思想和物理中的“粗粒化”思想，以及HP模型，根据a，t，c，g的化学结构分类，提出了DNA序列的特征序列概念（σ-，τ-，σ∩τ-）并推广到蛋白质序列中，从而给出一种数值刻划，将蛋白质序列简化成一个（0，1）序列，基于上述给出特征序列的方法，根据氨基酸分子量与简并度的关系，提出了另外一种DNA序列的特征序列概念（-）并推广到蛋白质序列中，进而给出了另外一种数值刻划，将蛋白质序列简化成一个（0，1，2）序列，通过比较RHD基因和RHCE基因的特征序列的数值刻划图，得出RHD基因和RHCE基因均偏爱使用低分子量且高简并度的氨基酸。  

Purification and characterization of a recombinant alpha-N-acetylgalactosaminidase from Clostridium perfringens

Clostridium perfringens alpha-N-acetylgalactosaminidase (alphaNAG) hydrolyzed the terminal N-acetyl-alpha-d-galactosamine from the blood type A(2) antigen producing H antigen, blood type O. Blood type O is universally compatible in the ABO system. Purification of the native enzyme is difficult with very low yields. To obtain the enzyme in satisfactory yield, the gene encoding the clostridial enzyme was cloned in an Escherichia coli T7 expression system. A highly purified preparation of recombinant alphaNAG was obtained from cell lysates by ion-exchange chromatography and high-pressure liquid chromatography. The final preparation was homogeneous by SDS-PAGE with a molecular mass of 71.96kDa and the native molecular weight of 72.42kDa. The enzyme was highly selective for terminal N-acetylgalactosamine residues. No other significant exoglycosidase activities, particularly neuraminidase, were detected. The pH optimum of the enzyme was between 6.5 and 7.0 and activity was relatively unaffected by ionic strength. ELISA experiments demonstrated activity against blood type A(2) epitope. These characteristics were similar to those of native alphaNAG from C. perfringens. With adequate expression in E. coli, sufficient recombinant alphaNAG enzyme mass can be obtained for potential use in enzymatic conversion of human blood type A(2) red blood cells to universally transfusable type O red blood cells.  

Identification,molecular cloning and expression of an alpha-N-acetylgalactosaminidase gene from Clostridium perfringens

The Clostridium perfringens gene encoding the previously characterized alpha-N-acetylgalactosaminidase (alphaNAG) was identified by protein microsequencing and database searching. The alphaNAG protein, designated AagA, was found to be encoded by a hypothetical gene of unknown function in the recently completed genome sequence of C. perfringens strain 13. The deduced translation product of 629 amino acid residues possessed a region of limited homology to several hypothetical open reading frames, an enterotoxin of unknown function and several known or predicted alpha-galactosidases, but did not exhibit homology to any of the multiple sequenced eukaryotic alphaNAG proteins. The C. perfringens aagA gene, encoding AagA, was cloned in an Escherichia coli T7 expression system, resulting in recombinants exhibiting high-level expression of the expected alphaNAG activity. To our knowledge, this is the first report of the cloning and expression of a bacterial alphaNAG-encoding gene and represents an important step in the development of recombinant alphaNAG as a tool in the enzymatic conversion of blood group antigens.  

The AB blood group system in wild felids

The blood type of 131 non-domesticated felids belonging to 26 felid species was surveyed in this study. Based upon a tube hemagglutination assay established for domestic cats, 80% of felids had type-A, 18% type-B, and 2% type-AB blood. Felids in the Puma group and African and Asian golden cats had blood type B, whereas all other species were found to have blood type A. Two cheetahs and one bobcat had type-AB blood. Red cell glycolipids analysed by high performance thin layer chromatography revealed a similar ganglioside pattern in wild cats as reported in domestic cats. Independent of the AB blood group system, incompatible blood crossmatch reactions were detected between different felid groups. In conclusion, wild felids display the same AB-erythrocyte antigens as domestic cats, and the same blood typing procedures can be applied for wild and domestic felids.  

Genetic variation within the Hereford breed of cattle

Genetic differentiation among Hereford populations from Britain, Ireland, Sweden, Canada and New Zealand together with six other beef breeds was assessed using blood type polymorphisms. Changes in the genetic structure of the British Hereford population over time were also examined. Loci surveyed were seven red cell antigen systems (A, B, C, F, L, S, Z), and two serum protein loci (transferrin and albumin). Within group variation was measured by the average expected heterozygosity, and between group relationships by genetic distance. There was significant genetic differentiation among Hereford populations from different countries. Differences between Hereford groups, however, were not as large as differences between breeds. There were also significance differences among British herds. The proportion of Canadian genes in the British 'hybrid' population was estimated to have increased from 0·42 (±0·34) in the 1970s to 0·98 (±0·11) in the 1990s. Canadian Hereford groups were found to be less heterozygous than other groups, and replacement of the British population with Canadian animals may lead to loss of variation. Breeding strategies that preserve original native genes in British Hereford populations should be considered by commercial breeders, in order to prevent the long-term loss of genetic variation within the breed.