釉原蛋白微损分析方法鉴定人骨性别的有效性与优化  

Verification and optimization of micro-destructive proteomics method for sex determination of archaeological human remains

作　　者：张宝帅 吴晓桐[1,2] 吴高 金正耀[1,2] 范安川 ZHANG Baoshuai;WU Xiaotong;WU Gao;JIN Zhengyao;FAN Anchuan(USTC Archaeometry Laboratory,University of Science and Technology of China,Hefei 230000;Department for the History of Science and Scientific Archaeology,University of Science and Technology of China,Hefei 230000;Core Facility Center for Life Sciences,University of Science and Technology of China,Hefei 230000)

机构地区：[1]中国科学技术大学科技考古实验室,合肥230000 [2]中国科学技术大学科技史与科技考古系,合肥230000 [3]中国科学技术大学生命科学实验中心,合肥230000

出　　处：《人类学学报》2023年第4期472-487,共16页Acta Anthropologica Sinica

基　　金：国家自然科学基金(41303080);中国科学院青年创新促进会会员人才项目(2018499);中国科学技术大学学术领军人才培养计划(2018)。

摘　　要：釉原蛋白微损分析方法是一种新的考古人类遗骸性别鉴定方法。最新的研究表明,该方法可以对未成年个体和保存较差的人类遗骸进行可靠的性别鉴定,而中国尚未开展相关研究。为验证和优化基于微损取样的釉原蛋白性别鉴定方法,本文对中国不同地区新石器时代至秦汉时期11个遗址的24个样品进行了研究。结果表明,基于该方法测试的所有样本都获得了可靠的性别信息。同时,本文对该方法的实验步骤和性别判定标准进行了优化:1)根据所获43组实验结果,提出以30个肽段作为排除男性假阴性可能的参考阈值;2)对盐酸溶液中的多肽进行多次提取是增加多肽数目和种类的有效途径,同时不会对样品造成二次破坏;3)数据检索过程中,Oxidation(M)和Deamidation(NQ)三种可变修饰对获得可靠的性别鉴定结果是必要的。Proteomics analysis of sex-specific amelogenin peptides in tooth enamel has a high accuracy in determining sex,which is especially useful for poorly preserved fossil human remains.Destructive sampling has been used in most studies,which consumed a considerable been used in most studies,so less destructive analysis such as the one described here is more desirable.Since this method was developed,many researchers have used it to identify sex of archaeological samples.To verify and optimize this approach,the use of micro-destructive analysis of teeth with different preservation conditions is investigated using 24 samples from 11 sites(from Neolithic to the Qin-Han period).This method obtained sex identification results consistent with physical anthropology in all 18 archaeological samples with known sex.At the same time,six individuals of unknown sex were reliably judged,which shows that the microdestructive analysis method has good applicability to tooth samples.We optimized this method in sampling and data analysis,noting the following points.1)There is a certain linear relationship between number of total peptides and Ay-specific peptides.Combined with the fitting curve,30 peptide fragments were identified as the reference threshold to exclude male false negative results.For samples without Ay-specific peptide segments,if the value was higher than 30,it was determined to be female;otherwise it’s sex was not determined.2)Multiple extraction of peptides from HCl solution is an effective way to increase number and types of peptides without secondary damage to teeth.3)Three kinds of variable modification(Oxidation(M)and Deamidation(NQ))are necessary in database search.At the same time,the relationship between different factors and protein content was also evaluated.At present,there is no relationship between burial or thermal age,distribution area and preservation status of amelogenin peptides.How to characterize the preservation status of amelogenin in samples by appearance or other means remains to be further stud

关 键 词：釉原蛋白 性别鉴定 微损 

分 类 号：Q987[生物学—遗传学]