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

采用电泳、等电聚焦、SDS-PAGE等技术对血清蛋白的遗传多态性进行了研究。分型样品采用蛋白酶（羧肽酶B、羧肽酶Y、木瓜蛋白酶、v8蛋白酶等）和唾液酸酶预处理。电泳后的图案通过电印迹或免疫固定可见。目前得到的结果如下。1. 我们在日本人群中发现了更多的C3新变体。通过比较研究，其中一种变体被命名为FO2，并且在FO2分子中发现了Arg到Gln的氨基酸取代。（提交出版）。2. 从血浆或血清样本中成功进行C4分型。样品在电泳前分别用羧肽酶B和唾液酸酶处理。C4多态性不仅对法医学领域的亲子鉴定有重要价值，对临床医学病因学研究也有重要意义。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沉默等位基因（C4**AQO）与这些疾病密切相关。(印刷中)。3. 在日本人的ATIII系统中发现了1个共有等位基因和4个变异等位基因。我们的报告（已出版）描述了打字方法、变体和命名法（字母数字代码）。4. 在补体HF的ief模式中，观察到许多非遗传控制的条带；需要进一步的研究。在补体BF和Cls分型过程中获得了一些新的发现（结果将在其他地方描述）。5. 对血迹样本进行遗传标记检测。红花叶提取物对HP和TF分型均有较好的效果。目前，我们正在研究用红花和酶处理提取物作为电泳样品，确定血迹的遗传标记。

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.