【作者】 张子波；

【导师】 杨康鹃；

【作者基本信息】 延边大学 ， 生物化学与分子生物学， 2003， 硕士

【摘要】 为了研究延边朝鲜族人群Ⅰ型胶原蛋白A2(COL1A2)基因MspⅠ多态性特点的存在和等位基因频率，并与其它地区人群进行比较，以探讨该群体的遗传结构及遗传背景，并为该群体的起源、迁移及与COL1A2基因相关性疾病的研究提供重要的遗传信息。在吉林省延边地区共收集朝鲜族血样120份（男：60份；女：60份），均为三代内无血缘关系，且为同一民族的健康个体。外周静脉血3ml，ACD抗凝，-80℃冻存至提取基因组DNA。按酚/氯仿抽提法提取基因组DNA ；用聚合酶链反应(PCR)方法，采用特定的PCR引物扩增目的基因，从人类外周血基因组DNA中分离到一个长约600bp的片段,采用Dynabeads固相单链分离直接测序法测定证明分离的片段是COL1A2基因；应用限制性片段长度多态性(RFLP)的技术方法，选用限制性内切酶MspⅠ进行酶切上述PCR扩增的目的基因DNA片段；琼脂糖凝胶电泳检测PCR产物及酶切产物；凝胶自动成像系统紫外线测仪上观察酶切结果，直接判读每个个体的基因型，计算基因型频率和等位基因频率。延边朝鲜族人群Ⅰ型胶原蛋白A2(COL1A2)基因存在多态性。120名朝鲜族中无MspⅠ酶切位点的纯合个体89例，基因型(P-P-)频率为74.17%，有MspⅠ酶切位点的杂合个体22例，基因型(P+P-)频率为18.33% ，有MspⅠ酶切位点的纯合个体9例，基因型(P+P+)频率为7.5% ； 240个COL1A2等位基因中MspⅠ酶切位点基因<WP=4>(P+)出现的频率为16.7%。小于苏联莫斯科（高加索人）俄罗斯人﹑贝加尔湖（蒙古人）的Burtay人，高于意大利人﹑挪威人﹑北美人﹑南非人群的好望角有色人种和黑人。显示了不同的种族人群的COL1A2基因MspⅠ多态性的基因频率是不同的，说明MspⅠ多态性特点可以作为延边朝鲜族的遗传标记。更多还原

【Abstract】 To study the polymorphism and allele frequency MSPⅠ site at COL1A2 gene for koreans Yanbian ,and contrast to other area population ,and give important genetic information to understand the origin and moving and correlate disease at COL1A2 gene of the population and genetic structure and genetic background; and to provid part medical genetics data for creation of immortalization cell bank for koreans China.we extracted human genomic DNA using phenol/chloroform extracting; and detection polymorphism MSPⅠsite at COL1A2 gene by the PCR -RFLP technique. A 600bp fragment from human genomic DNA was amplied by the PCR -RFLP technique,The PCR product was partly sequenced with dynabeads-streptavidin solid ,to detect PCR product and enzyme cut product by agovid gel electrophoresis and observing result in infared meter gel aut made aerial image ,Direct reading genotypic and calculation genotypic frequency and gene frequency.In 120 coreans, homozygotes with no digestion MSPⅠsite are 89 and genotype frequency (P-P-) is 74.17% ; heterozygotes with digestion MSPⅠsite are 22 and genotype frequency (P+P-) is 18.33% ; homozygotes with digestion MSPⅠsite are 9 and (P+P+) is 7.5% . The frequency of P+ is 16.7% in 240 allele of COL1A2 gene 。The polymorphism site at COL1A2 gene is associated with<WP=6>coreans Yanbian ;and distribute of the polymorphism MSPⅠ site at COL1A2 gene is different in coreans Yanbian and other areas, so the polymorphism MSPⅠsite of COL1A2 gene as genetic marker of koreans Yanbian .更多还原