Development of molecular sex determination methods and their application to archaeological material sourced from the Raymond Dart collection of human skeletons

摘要

Within biological anthropology there is a movement toward the use of mo-udlecular methods and their application to anthropological questions. In SouthudAfrica there is great potential for research in anthropological genetics as thisudcountry is a vast repository of skeletal remains. This study aims to developudnovel methods of molecular sex determination for skeletal material and applyudthese to an archaeological collection of historical importance.udThe bone extraction method developed here is minimally destructive to skeletaludmaterial and does not interfere with any anthropometric landmarks. The ex-udtracted bone powder is subjected to DNA extraction procedures adapted forudbone. In addition, two novel systems of molecular sex determination ideal forudskeletal material are developed on the amelogenin gene, beginning in intronud2-3, spanning exon 3 and ending in intron 3-4. This area is optimal for sex-uding, as it includes 14 sex specific polymorphic regions in addition to an indelud(insertion or deletion of nucleotides). Once these procedures of molecular sexuddetermination are optimised and working with 100% efficiency on the controls,udthey will be applied to a collection of miscellaneous archaeological skeletonsud(ex-situ) sourced from the Raymond Dart Collection of Human Skeletons (DartudCollection). This collection is used to optimise these techniques for specimens derived from an archaeological context. These methods yielded 46.66% sex res-udults for the ex-situ sample, which is within the normal range for ancient DNAudstudies. These novel methods are then applied to an archaeological sample withudgood provenance, this being 36 skeletons of Chinese indentured miners sourcedudfrom the Dart Collection. While a previous morphometric study showed thatudthis collection was represented by equal male to female ratios, the historical records suggest however that very few women accompanied these Chinese la-udbourers. In using these procedures, 41.93% of this sample produced results alludof which were male, correlating well with the historical records. The value ofudmolecular approaches to investigate sex determination is that they do not relyudon intact specimens as morphmetric methods do. Therefore in cases where theuduse of morphometrics is complicated, molecular approaches offer an accurateudsolution for diagnosing sex.udThe techniques of bone and DNA extraction are applicable for both humanudand animal skeletal tissue, and the methods of molecular sex determinationudare optimal for archaeological or forensic derived human skeletal material.