中文摘要：

背景家庭血胆脂醇过多(FH ) 是与低密度脂蛋白(LDL ) 铺平导致早熟的冠的心疾病(CHD ) 的提高的血浆联系的正染色体的混乱。由于长期的 hyperlipemia， FH 病人将介绍 endarterium 变厚和动脉粥样硬化。在现在的学习，我们为可能的变化扫描了一个临床上诊断的正染色体的基因血胆脂醇过多家庭的相关基因并且建立了 proprotein convertase subtilisin/kexin 类型 9 的变化( PCSK9 )的真核细胞的表示向量有基因再结合的基因在低密度脂蛋白受体上调查变化的贡献的技术( LDL-R )新陈代谢和功能 alternation.Methods 变化察觉为 LDL-R 被进行， apolipoprotein B100 ( apoB10 野类型 PCSK9 基因(WT-PCSK9 ) 的全身的 cDNA 从 Bel-7402 被获得。地点 mutagenesis 被用来真核细胞的表示向量带建立 recombinant PCSK9 基因和插入的碎片的病原的类型被定序。与是的空白的向量控制， liposome transfection 方法习惯于 transfect 有 recombinant plasmid 的 Bel-7402 房间。LDL-R mRNA 的表示被 RT-PCR 检验。PCSK9 和 LDL-R 蛋白质的表示被西方的弄污决定。结果在 PCSK9 基因的 918 核苷酸的 GT 变化在 codon 由丝氨酸导致了精氨酸的替换 306 exon 6。在定序以后，确定的表示向量的插入的碎片有正确尺寸和顺序，异种高度在 Bel-7402 房间被表示，这被证实。在 LDL-R mRNA 的层次没有重要变化。在房间是由 WT-PCSK9 plasmid 的 transfected 以后， LDL-R 成熟蛋白质被 57% 减少。在房间是由变异的 R306S 的 transfected 以后，成熟 LDL-R 是由灰色的规模扫描证实了被12%显著地减少，建议新变异的 R306S 罐头显著地减少成熟 LDL-R protein.Conclusions 的表示 PCSK9 基因的一个新奇错误感觉变化， R306S ，被发现并且真核细胞的表示向量 PCSK9 变异、野类型基因被建立。在 LDL-R mRNA 的层次没有重要变化。R306S 变化能显著地导致 LDL-R 成熟蛋白质表示的减少，它可能是 FH 家?

英文摘要：

Background Familial hypercholesterolemia （FH） is an autosomal disorder associated with elevated plasma low density lipoprotein （LDL） levels leading to premature coronary heart disease （CHD）. As a result of long-term hyperlipemia, FH patients will present endarterium thickening and atherosclerosis. In the present study we scanned the related gene of a clinically diagnosed autosomal genetic hypercholesterolemia family for the possible mutations and established eukaryotic expression vector of mutation of proprotein convertase subtilisin/kexin type 9 （PCSK9） gene with gene recombination technique to investigate the contributions of the variation on low density lipoprotein receptor （LDL-R） metabolism and function alternation.Methods Mutation detection was conducted for LDL-R, apolipoprotein B100 （apoB100） and PCSK9 gene with nucleotide sequencing in a Chinese FH family. The full-length cDNA of wild type PCSK9 gene （WT-PCSK9） was obtained from Bel-7402. Site mutagenesis was used to establish the recombinant eukaryotic expression vector carrying pathogenic type of PCSK9 gene and the inserted fragment was sequenced. With the blank vector as control, liposome transfection method was used to transfect the Bel-7402 cells with recombinant plasmid. The expression of LDL-R mRNA was examined by RT-PCR. PCSK9 and the expression of LDL-R protein were determined by Western blotting. Results The G→T mutation at the 918 nucleotide of PCSK9 gene resulted in the substitution of the arginine by a serine at the codon 306 of exon 6. After sequencing, it was confirmed that the inserted fragment of established expression vector had correct size and sequence and the mutant was highly expressed in Bel-7402 cells. There was no significant variation in the levels of LDL-R mRNA. LDL-R mature protein was decreased by 57% after the cells were transfected by WT-PCSK9 plasmid. Mature LDL-R was significantly decreased by 12% after the cells were transfected by R306S mutant as evidenced by gray scale scanning, suggesting