Study on the Genetic Polymorpphisms of Serum Proteins from Protease-treated Samples and its Application to Forensic Medicine.

采用电泳法、等电聚焦法和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法(SDS-PAGE)研究了血清蛋白的遗传多态性。对分型样品进行了蛋白酶(羧基肽酶B、羧基肽酶Y、木瓜蛋白酶、V8-蛋白酶等)处理。和唾液酸酶。用电印迹或免疫固定的方法显示电泳后的图谱。到目前为止，取得的结果如下。1.我们在日本人的化蛹中发现了C3的其他新变种。通过比较研究发现，一个变异体被命名为FO2，并在FO2分子中发现了Arg到Gln的氨基酸取代。(已呈交以供发表)。2.从血浆或血清标本中成功地进行了C4配型。样品在电泳前用羧基肽酶B和唾液酸酶处理。C4基因多态性对法医学亲子鉴定和临床病因学研究具有重要意义。我们检测了IgA肾病、天疱疮、大疱性类天疱疮或系统性红斑狼疮患者的样本，发现Alocus的沉默等位基因(C4**AQO)与这些疾病密切相关。3.在日本人中发现了ATIII系统的1个常见等位基因和4个变异等位基因。我们的报告(在印刷中)描述了分型方法、变体和命名(字母-数字代码)。4.在补体Hf的IEF带型中，观察到了许多非遗传控制的条带，有待进一步研究。在补体BF和CLS的分型过程中，已经获得了一些新的发现(结果将在其他地方描述)。5.对血痕标本进行了遗传标记检测。用弗洛里硅胶处理血迹提取液对HP和Tf分型有较好的效果。我们正在研究用花瓣提取液和酶滴定提取液作为电泳法测定血痕的遗传标记。

We studied the genetic polymorphisms of serum proteins by using electrophoresis, isoelectric focusing and SDS-PAGE.Samples for the typings were pre-treated with protease(carboxypeptidase B, carboxypeptidase Y, papain, V8-protease, etc.) and sialidase. Patterns after electrophoresis were visualized by electroblotting or immunofixation. The results obtained so far are as follows. 1. We found additional new variants of C3 in a Japanese pupulation. One variant was named as FO2 from a comparative study, and an amino acid substitution of Arg to Gln was found in the FO2 molecule. (submitted for publication). 2. C4 typing was performed successfully from plasma or serum samples. The samples were treated with both carboxypeptidase B and sialidase prior to electrophoresis. The C4 polymorphism is invaluable not only for the paternity test in medico-legal field but also for the study of etiology in clinical medicine. We examined the samples from patients with IgA-nephropathy, pemphigus, bullous pemphigoid or systemic lupus erythematosus, and found that the silent allele of Alocus(C4**AQO) was strongly associated with those diseases.(in press). 3. We found one common and four variant alleles of ATIII system in Japanese. Typing method, variants and nomenclature(alpha-numeric code) were described in our report (in press). 4. In the IEF-patterns of complement HF, many non-genetically controlled bands were observed ; the further study is required. Severa new findings have been obtained during the typings of complement BF and Cls (the results will be describled elsewhere). 5. Detection of genetic marker has been performed from bloodstain sample. The treatment of bloodstain extracts with florisil gives good results for HP and TF typings. We are now studying the determination of genetic markers from bloodstains by using the florisil-and enzyme-trated extracts as the samples of electrophoresis.