目的通过小样本外显子测序与公用数据库比对进行生物信息挖掘,探寻汉族人群高原红细胞增多症(high altitude polycythemia,HAPC)变异与发病的相关性。方法纳入汉族高原红细胞增多症男性患者4例和高原健康男性5例,采集静脉血并提取DNA进...目的通过小样本外显子测序与公用数据库比对进行生物信息挖掘,探寻汉族人群高原红细胞增多症(high altitude polycythemia,HAPC)变异与发病的相关性。方法纳入汉族高原红细胞增多症男性患者4例和高原健康男性5例,采集静脉血并提取DNA进行全外显子组测序;将测序数据进行功能学富集分析(gene ontology,GO&kyoto encyclopedia of genes and genomes,KEGG)构建突变基因蛋白质互作网络(protein protein interaction,PPI),选择显著性最高的基因,筛选潜在关键HAPC相关变异位点,初步探究相关变异与HAPC发病可能的机制。结果通过全外显子测序,发现HAPC相关的突变基因216个,其中表皮生长因子(epidemal growth farctor,EGF)基因在功能学富集分析(GO&KEGG),蛋白质互作网络(PPI)中具有高度显著性,初步筛选出5个潜在关键HAPC相关变异位点,通过生物信息学预测及分析,最终筛选出EGF基因上3个可能的变异c.2124G>A(p.Met708Ile)、c.2351A>T(p.Asp784Val)以及c.2759A>T(p.Glu920Val),可能是HAPC的突变位点,与HAPC发病相关。结论本研究表明汉族EGF基因的变异与HAPC发病存在相关性。EGF基因的变异可能通过干扰RNA剪接,改变RNA二级结构,影响蛋白质结构,进而影响EGF的生成及HAPC的发生发展。展开更多
The epidermal growth factor （EGF） has been shown to promote the proliferation of various types of cells, to maintain the physiological function of the mucosa of the digestive tract, and to promote the healing of the...The epidermal growth factor （EGF） has been shown to promote the proliferation of various types of cells, to maintain the physiological function of the mucosa of the digestive tract, and to promote the healing of the gastric and duodenal ulcers. It has been expressed in many types of bacteria and yeasts. In this article, a bio-reactor was constructed, namely, the human EGF （hEGF） transgenic mini-tomato. On the basis of hEGF gene sequence, a tomato codon preference hEGF gene was chemically synthesized, and it was constructed into the plant expression vector pCAMBIA2300. The transgenic tomato plants containing gene hEGF were obtained through Agrobacterium-mediated transformation. The expression product was determined by radioimmunoassay （RIA） and showed a yield of 3.48± 1.01 ng/g fresh fruits. The intragastric gavage （ig） administration of the rhEGF-containing juice of the transgenic tomato （equivalent to 24 ng rhEGF per mouse a day） for 15 days could significantly protect mice against the alcohol-induced ulceration. The ulcer index, expressed as a degree of the stomach lesion, decreased from 42.20 ±18.13 to 16.25 ±9.57.展开更多
文摘目的通过小样本外显子测序与公用数据库比对进行生物信息挖掘,探寻汉族人群高原红细胞增多症(high altitude polycythemia,HAPC)变异与发病的相关性。方法纳入汉族高原红细胞增多症男性患者4例和高原健康男性5例,采集静脉血并提取DNA进行全外显子组测序;将测序数据进行功能学富集分析(gene ontology,GO&kyoto encyclopedia of genes and genomes,KEGG)构建突变基因蛋白质互作网络(protein protein interaction,PPI),选择显著性最高的基因,筛选潜在关键HAPC相关变异位点,初步探究相关变异与HAPC发病可能的机制。结果通过全外显子测序,发现HAPC相关的突变基因216个,其中表皮生长因子(epidemal growth farctor,EGF)基因在功能学富集分析(GO&KEGG),蛋白质互作网络(PPI)中具有高度显著性,初步筛选出5个潜在关键HAPC相关变异位点,通过生物信息学预测及分析,最终筛选出EGF基因上3个可能的变异c.2124G>A(p.Met708Ile)、c.2351A>T(p.Asp784Val)以及c.2759A>T(p.Glu920Val),可能是HAPC的突变位点,与HAPC发病相关。结论本研究表明汉族EGF基因的变异与HAPC发病存在相关性。EGF基因的变异可能通过干扰RNA剪接,改变RNA二级结构,影响蛋白质结构,进而影响EGF的生成及HAPC的发生发展。
基金This work was supported by the Innovation Group Development Project of the Ministry of Education of China (No. IRT0435).
文摘The epidermal growth factor （EGF） has been shown to promote the proliferation of various types of cells, to maintain the physiological function of the mucosa of the digestive tract, and to promote the healing of the gastric and duodenal ulcers. It has been expressed in many types of bacteria and yeasts. In this article, a bio-reactor was constructed, namely, the human EGF （hEGF） transgenic mini-tomato. On the basis of hEGF gene sequence, a tomato codon preference hEGF gene was chemically synthesized, and it was constructed into the plant expression vector pCAMBIA2300. The transgenic tomato plants containing gene hEGF were obtained through Agrobacterium-mediated transformation. The expression product was determined by radioimmunoassay （RIA） and showed a yield of 3.48± 1.01 ng/g fresh fruits. The intragastric gavage （ig） administration of the rhEGF-containing juice of the transgenic tomato （equivalent to 24 ng rhEGF per mouse a day） for 15 days could significantly protect mice against the alcohol-induced ulceration. The ulcer index, expressed as a degree of the stomach lesion, decreased from 42.20 ±18.13 to 16.25 ±9.57.
