延边朝鲜族低密度脂蛋白受体基因PvuⅡ多态位点的研究

Detection of the polymorphic PvuⅡ site at LDL receptor gene for Koreans-Chinese in Yanbian

[目的 ]研究延边朝鲜族人群低密度脂蛋白受体基因内含子 15PvuⅡ多态性位点的存在和等位基因频率 ,以探讨该群体的遗传结构及遗传背景 .[方法 ]按酚 /氯仿抽提法提取基因组DNA ;用聚合酶链反应 限制性片段长度多态性技术从人类外周血基因组DNA中分离到一个长约 810bp的片段 ,采用Dynabeads固相单链分离直接测序法测定 ,证明分离的片段是否是低密度脂蛋白受体基因内含子 15 ;用限制性内切酶PvuⅡ酶进行酶切 ,进行琼脂糖凝胶电泳 ,在凝胶自动成像系统紫外线检测仪上观察酶切结果 .[结果 ]延边朝鲜族人群低密度脂蛋白受体基因存在PvuⅡ多态性 .12 0名朝鲜族中无酶切位点的纯合个体 92例 ,基因型 (P-P-)频率为 76 6 % ;有酶切位点的杂合个体 2 0例 ,基因型 (P+ P-)频率为16 7% ;有酶切位点的纯合个体 8例 ,基因型 (P+ P+ )频率为 6 7% .2 4 0个低密度脂蛋白受体等位基因中PvuⅡ酶切位点基因 (P+ )的出现频率为 15 % .[结论 ]延边朝鲜族人群低密度脂蛋白受体基因也存在PvuⅡ多态性位点 ,说明PvuⅡ多态性特点可以作为延边朝鲜族的遗传标记 .

OBJECTIVE?To study the polymorphism and allele frequency of pvuⅡ site intron 15 at LDL receptor gene and genetic structure and background for Koreans Chinese in Yanbian.?METHODS? Humangenomic DNA was extracted by using phenol/chloroform extracting.A 810?bp fragment from human genomic DNA was amplied by the PCR RFLP technique, the PCR product was partly sequenced with dynabeads streptavidinsolid phase sequencing technique and directly analysized by digestion of pvuⅡ,and agarose gel electrophoresis, and detected results of digestioned in Gel Doc 1000(Bio Rad).?RESULTS?PvuⅡ polymorphism intron 15 at the LDL receptor gene is associated with Koreans Chinese in Yanbian,in 120 Koreans Chinese, homozygotes with no digestion site are 92 and genotype frequency (P -P -) is 76 6% ; heterozygotes with digestion site are 20 and genotype frequency (P +P -) is 16 7% ; homozygotes with digestion site are 8 and genotype frequency (P +P +) is 6 7%.The frequency of P + allele (presence of pvuⅡ site) is 15% in 240 allele of LDLR genes.?CONCLUSION?PvuⅡ polymorphism at intron 15 of LDLR genes is associated with Koreans Chinese in Yanbian yet, so pvuⅡ polymorphism site as genetic marker of Koreans Chinese in Yanbian.