用快速高效液相色谱系统分离纯化Gc蛋白

报告Gc蛋白的一种分离方法，为免疫制备抗Gc血清制备抗原。硫酸该分级沉淀出含Gc的人血清组份，BlueSepharoseCL－6B亲和色谱除去含Gc硫酸铸分级沉淀的人血清组份中的白蛋白，快速高效液相色谱系统过Alkyl-SuperoseTMHR5／5疏水色谱柱和MonoQHR5／5离子交换柱。聚丙烯酸胺凝胶解离、非解离电泳证明，Gc蛋白得到了彻底纯化。免疫固定显示：纯化出的蛋白确实是Gc蛋白．为1-1型。快速高效液相色谱为蛋白质的纯化提供了新的技术及仪器手段。     

Gc亚型检测的复合MS-PCR法及其应用

目的探讨建立Gc亚型检测的复合MS-PCR法及其应用价值。方法根据Gc基因中的2处点突变,设计2对片段相差5bp的等位基因特异性引物和1条公共引物进行复合MS-PCR,分析Gc多态性,并调查武汉地区218例汉族无关个体Gc多态性和鉴定10例亲子关系。结果复合MS-PCR检测的Gc基因型,与AmpliTypePM试剂盒的分型结果一致;武汉地区汉族人群Gc基因的3个常见等位基因Gc1F、Gc1S、Gc2的基因频率分别为0.4816、0.2592、0.2592,观察杂合度(Hobs)、期望杂合度(Hexp)、多态性信息含量(PIC)、个人识别能力(DP)、非父排除率(PE)分别为0.6193、0.6359、0.6253、0.7974、0.3480,基因型分布符合Hardy-Weinberg平衡;真三联体和非真三联体亲子鉴定各5例,前者不排除父子关系,与常规STR分型一致,后者经Gc-MS-PCR分型排除2例。结论建立复合MS-PCR法检测Gc亚型在法医物证鉴定中有实用价值。     

人血清Hp、Tf、Gc型的同步检测

人血清Hp、Tf、Gc蛋白是在法医学个人识别及亲子鉴定中应用最为广泛的三种遗传多态性蛋白标记。[1][2][3]本文采用聚丙烯酰胺垂直板状凝胶电泳同时进行Hp、Tf、Gc三种蛋白的分型。经氨基黑、联苯胺分别染色后。Hp、Tf、Gc的电泳图谱清晰、重复性好,整个操作过程仅需6小时。作者还用该方法检测了上海地区人群Hp、Tf、Gc的分布频率。     

用垂直板状电泳同时测定Gc及Tf的多态性

型特异性组分(Gc)和转铁蛋白(Tf)都是人血清中的蛋白成份,具有遗传多态性.Smith(1957年)和Hirschfeld(1959年)分别用淀粉凝胶电泳和免疫电泳检测了Tf和Gc的多态性.Tf分为三种常见表型:TfBC、TfCC、TfDC.Gc又分为三种常见表型:Gc~(1-1)、Gc~(2-1)、Gc~(2-2).以后国内外许多学者对Tf及Gc的生化、遗传和免疫特征又做了大量的研究,并且测定了世界上不     

成都地区汉族Gc亚型的分布及血痕中Gc亚型的检测

作者用免疫固定薄层聚丙烯酰胺凝胶等电聚焦(PAGIF)技术,调查了成都地区无关的125名健康汉族人血清Gc亚型分布。其6种亚型频率(%)分别为:Ge1F=20.8,Ge1S=8.0,Gc1F-1S=18.4,Gc2-1F=30.4,Gc2-1S=16.0和Gc2=6.4。Gc的基因频率为:Cc~(1F)=0.452,Gc~(1S)=0.252和Gc~2=0.296。对保存于室温条件下20周的陈旧血痕进行了Gc亚型定型,获得满意结果。     

在Gc电泳图谱上读Hp表型

前言血型特异性成分(Gc)和结合珠蛋白(Hp)是两种血清球蛋白,分别具有三种遗传表型Gc1—1,Gc2-1,Gc 2—2,和Hp1-1,Hp2-1,Hp2-2,按孟德尔遗传定律遗传,是法医学个人识别及亲权鉴定的两个重要遗传标记。一般情况下,检测血清Gc和Hp需要分别配制凝胶、凝胶缓冲     

广州地区人群血清Gc型的测定——聚丙烯胺酰凝胶圆盘电泳法

<正> 型特性异成分(Group Specific Component,简称Gc)是人血清中的一组α_2球蛋白。1959年,Hirschfeld用免疫电泳法首先发现Gc,并将人群分为Gc1-1,2-1,和2-2三型。此后,许多学者对Gc进行了多方面的研究,证实Gc的多态性是由按孟德尔遗传方式的基因控制的。许多国家和地区的法医学工作者已把Gc分型应用于实际检案工作中。     

应用窄范围两性电解质聚丙烯酰胺凝胶等电聚焦法检验人血痕中的Gc亚型

采用窄范围两性电解质聚丙烯酰胺凝胶等电聚焦加免疫吸印技术,对人血痕中 Gc 亚型进行分型,并调查了北京地区284名无关汉族群体的 Gc 亚型的分布及基因频率。结果显示:Gc~(IF)0.4287,Gc~(IS)0.2500.Gc~20.3222.观察值与期望值吻合度良好(ΣX~2=0.7530,P>0.80).Gc 的个人识别率为0.6507.且室温下保存7周的血痕可以准确判型。将调查结果与中国其它地区及国外不同人种 Gc 亚型的表型的分布与基因频率进行了比较,发现地理及人种间差异明显。     

用免疫电泳技术检测人血痕中Gc的表型

<正> 在60年代初,有人尝试用免疫电泳技术对人血痕进行 Gc 定型,均以血痕中的Gc 蛋白在电泳时,向阳极飘移而告失败。Shinomiya 和 Baelen 发现 Gc 蛋白飘移的原因是与其血痕中破碎细胞释放出来的肌动蛋白结合形成复合物所致。用4M 盐酸胍处理血痕,可使复合物解聚,恢复 Gc 的正常电泳迁移率。     

型特异性成分(Gc,维生素D结合蛋白)型的研究现状

<正> 型特异性成分(Group specific component,Gc)是一组存在于血清中的生物活性相同,结构相似的α_2球蛋白。自1959年Hirschfeld首次报道Gc蛋白的多态性以来,学者们对Gc的化学组成、生物学功能、遗传特征及遗传规律进行了深入的研究。Gc作为一种遗传标记,在国外已广泛地应用于法医学个人识别和亲子鉴定。为了进一步在我国扩广应用,现综述介绍如下。     

Vitamin D binding protein (Gc) subtypes by isoelectric focusing and immunoblotting

The polymorphism of the human vitamin D binding protein (Gc system) was investigated in a total of 149 sera from unrelated healthy Egyptians residing in Tanta City, Gharbiya Governorate, Nile Delta of Egypt, using isoelectric focusing (IEF) in thin-layer polyacrylamide gel followed by immunoblotting. The estimated gene frequencies were Gc1s = 0.540, Gc1f = 0.242 and Gc2 = 0.218.     

The typing of group-specific component (Gc protein) in human blood stains

It is known that the typing of group-specific component (Gc protein) in human blood stains is difficult since Gc protein of the extracts of blood stains migrates more anodally to the α₁-globulin region in agar-gel immunoelectrophoresis, while Gc protein in liquid blood normally migrates to the α₂-globulin region. We have reported that the Gc protein found in the α₁-region is the result of binding of actin to Gc protein (Shinomiya, K., Kimura, H., Yoshida, K., and Shinomiya, T., J. Biochem., 92 (1982) 1163–1171, which renders it difficult to determine the Gc-phenotypes in the blood stains. On the basis of the above findings, we developed the method of phenotyping the Gc protein of human blood stains by agar-gel immunoelectrophoresis. Since the binding activity of actin to Gc protein is lost after treatment with a high concentration of guanidine HCl, the extracts of blood stains were treated with 4 M guanidine HCl to dissociate Gc protein and actin and then dialyzed to remove guanidine HCl. By this method we are able to determine the phenotypes of Gc protein in blood stains. The method we have developed is a useful tool in the forensic laboratory.     

Rapid phenotyping of the group specific component by immunofixation on cellulose acetate

Determination of the genetically controlled variants of the polymorphic Gc system was achieved by electrophoresis on cellulose acetate membranes followed by immunofixation with a specific anti-Gc antiserum. The method is applicable to plasma, whole hemolyzed blood, and dried blood. Multiple specimens can be analyzed simultaneously within 60 to 80 min. The cellulose acetate electrophoretogram of the Gc variants remains as a permanent record.     

Gc subtypes determined by ultrathin-layer isoelectric focusing. Distribution in the Veneto population (Italy)

The distribution of Gc phenotypes in the population of Veneto was investigated by ultrathin-layer isoelectric focusing. In our sample (n = 732) the six common phenotypes, Gc 1S, 1F, 1S1F, 2, 2-1S, 2-1F and a further phenotype, GC 1S1C3, were observed and the following frequencies calculated: Gc 1S = 0.560792; GC 1F = 0.159153; Gc2 = 0.277323; Gc 1C3 = 0.002732. Our gene frequencies have been compared with those found in other populations.     

Group specific component subtyping in bloodstains by separator isoelectric focusing in micro-ultrathin polyacrylamide gels followed by immunoblotting

The identification of group specific component (Gc) subtypes derived from blood-stains by separator isoelectric focusing in micro-ultrathin polyacrylamide gels (interelectrode distance: 50 mm) containing 4.5 to 5.4 pharmalytes is described. The separation achieved between Gc 1F and Gc 1S bands is compared favorably with that obtained using separator isoelectric focusing in conventional polyacrylamide gels dimensions (interelectrode distance: 110 to 120 mm). The technique is rapid and economical, and the immunoblotting method described is more sensitive than immunofixation followed by silver staining.     

A method for subtyping group-specific component in bloodstains

A method is described for subtyping group-specific component (Gc) derived from human bloodstains. Bloodstained cuttings were extracted in 6 M urea. The extracts were subjected to ultrathin-layer polyacrylamide gel isoelectric focusing in the pH 4.5-5.4 range. After isoelectric focusing, Gc was detected by immunofixation in cellulose acetate membranes. This method permitted the successful typing of Gc in at least four-month-old bloodstains maintained at room temperature. Bloodstains from 266 liquid blood samples of known origin were subjected to both this method and immunofixation conventional agarose gel electrophoresis with no phenotypic discrepancies observed. The Gc population data for Whites from Baltimore, Maryland, were homogeneous with white sample populations from other geographical locations within the U.S.A.; while Gc data from northern U.S.A. black sample populations appeared to be heterogeneous compared with a southern United States black sample population.     

The phenotypic frequencies of group specific component and alpha-2-HS-glycoprotein in three ethnic groups. The use of these proteins as racial markers in forensic biology

The phenotypic frequencies of group-specific component (Gc) and alpha-2-HS-glycoprotein (A2HS) were determined in White European, Asian and Afro-Caribbean populations. Typical allele frequencies were observed for Gc, with Gc 1S being the major allele for the first two groups and Gc 1F being the major allele for Afro-Caribbeans. For all groups the dominant A2HS allele was A2HS 1, although Asians had a significantly higher proportion of this allele than the White Europeans. Gc and A2HS either singly or in combination with other blood grouping systems provide good discriminating potential. The A2HS 10 allele was detected with a very low frequency in the White European group (A2HS 10 = 0.0013) and was not detected in the Asian group, while the Afro-Caribbean group had a relatively high frequency of this allele (A2HS 10 = 0.0966). The different distribution of the Gc 1F and A2HS 10 alleles in White Europeans and Asians compared with Afro-Caribbeans, can be used to determine the likelihood of blood coming from an Afro-Caribbean.