原发性下肢深静脉瓣膜功能不全的大隐静脉曲张致病相关基因的筛选与克隆

Screening and cloning of genes related to varicose great saphenous vein accompanying with primary deep vein velve insufficiency

目的 筛选和克隆原发性下肢深静脉瓣膜功能不全的大隐静脉曲张的致病相关基因。方法 利用mRNA荧光差异显示技术比较原发性下肢深静脉瓣膜功能不全患者的曲张大隐静脉隐 股瓣膜区组织中mRNA表达与对照组的差异 ,获得的差异表达cDNA片段经Northernblot验证后进行克隆和测序以探明其基因来源。 结果 共获得 37条差异表达的cDNA条带 ,经杂交验证其中 30条为阳性 (阳性率 80 %)。克隆及测序结果显示C610 片段 (NO 18克隆 )与麦 考综合征 (Mckusick Kaufmansyndrome,MKKS)基因的mRNA序列有 96 %的同源性。 结论 C610 片段 (NO 18克隆 )与麦 考综合征基因高度同源 ,与下肢大隐静脉曲张的发病密切相关。

Objective To screen the genes related to the occurrence and development of varicosis of the great saphenous vein in the patients with primary deep vein valve insufficiency. Methods Using mRNA fluorescent differential display reverse transcriptive polymerase chain reaction (FDD-RTPCR), different genes expressed in the varicose great saphenous veins in patients with primary deep vein valve insufficiency and corresponding normal human tissues were compared. Differentially expressed cDNA fragments confirmed by Northem blot were compared and then cloned into the pGEM-Teasy vector. Positive clones were selected and sequenced. All the sequences were put into Genebank and analyzed by BLASTN software to search for their genetic origins. Results Altogether 37 different cDNA fragments were obtained and 30 of which were confirmed by Northern blot. Analysis of the sequences by BLASTN software showed that C 610 fragment (NO.18 cDNA clone) shared 96% homology with the mRNA sequence of the human Mckusick-Kaufman syndrome gene (MKKS gene). Conclusion C 610 fragment is highly homologous with the mRNA sequence of the human MKKS gene and is closely related to the development of varicosis of the great saphenous vein in patients with primary deep vein valve insufficiency.