【摘要】 从条纹斑竹鲨中克隆肌钙蛋白基因,在大肠杆菌中进行表达,并检测其抑制血管生长的活性。文献发现人肌钙蛋白(TnI)具有抑制肿瘤血管生成的功能,根据人TnI的基因序列在条纹斑竹鲨鱼肝脏中克隆出于鲨鱼TnI基因,在大肠杆菌中进行表达,获得重组蛋白,通过MTT实验和鸡胚尿囊膜(CAM)实验验证其抑制血管生长的活性。条纹斑竹鲨TnI的cDNA全长692 bp,其中开放阅读框552 bp(编码具有183个氨基酸多肽)。该蛋白与角鲨鲨鱼TnI的氨基酸同源性最高达85.2%,而与人类、鲑鱼、斑马鱼的TnI的氨基酸同源性分别为68.9%,64.5%和62.3%。在大肠杆菌中进行重组表达,纯化获得蛋白,MTT实验证明其具有抑制血管内皮细胞生长的功能,鸡胚尿囊膜实验显示其有抑制血管生长的功能。结论:条纹斑竹鲨TnI具有抑制血管生成的功能。更多还原

【Abstract】 To clone the shark Troponin I(TnI) from whitespotted bambooshark(Chiloscyllium plagiosum Bonnet) and to express in E.coli.as well as to identify the anti-angiogenesis ability of this protein,the gene of human Troponin I(TnI) homolog from liver of whitespotted bambooshark(Chiloscyllium plagiosum Bonnet) was cloned,followed by recombinant expression in E.coli.Then the inhibitory ability of shark TnI for angiogenesis was investigated by MTT method and chicken embryo allantoic membrane(CAM) assessment.Results showed that the cDNA of shark TnI was 692 bp in length,with an ORF of 552 bp(encoding a polypeptide of 183 amino acids).It has the highest amino acid identity(85.2%) with dogfish shark TnI,whereas 68.9%,64.5% and 62.3% with TnIs of human,salmon and zebrafish,respectively.The recombinant shark TnI was found to inhibit the proliferation of human umbilical vein endothelial cells(HUVECs) in a dose-dependent fashion and angiogensis of chicken embryo allantoic membrane.The TnI of whitespotted bambooshark(Chiloscyllium plagiosum Bonnet) was confirmed to inhibit angiogenesis.更多还原