赤魟尾刺cDNA表达文库的构建

目的　以赤尾刺为出发材料,构建以真核表达载体pcDNA 3.0为基础的赤尾刺cDNA表达文库。方法　应用SMART^TM cDNA Library Construction 技术和初步生物信息学分析等方法。结果　通过对亚文库克隆的序列测定和初步生物信息学分析,已获得了56个赤新EST序列,其中已确定的全长cDNA克隆有24个,包括IPL肿瘤抑制因子、亲环素cyclophilin A、硒蛋白selenoprotein W、半胱氨酸蛋白酶抑制剂cystatin A、tyrosyl-tRNA synthetase 和 calcineurin 类似物蛋白等。结论　这些基因在赤中绝大多数均为首次发现。赤尾刺cDNA表达文库的成功构建,将有利于对赤尾刺中药应用的分子作用机理的深入研究

CONSTRUCTION OF cDNA EXPRESSION LIBRARY FROM THE CAUDAL SPINE OF DASYATIS AKAJEI

AIM A cDNA expressional library from the caudal spine of Dasyatis akajei was constructed into pcDNA 3 0 eukaryotical expression plasmid. METHODS SMART TM protocol was used for cDNA Library Construction and bioinformatics analysis was carried out. RESULTS Fifty six novel EST clones were obtained from the library, in which there were 24 full length clones, including IPL tumor suppressor gene, cyclophilin A, selenoprotein W, cystatin A, tryosyl tRNA synthetase and calcineurin homologous protein. CONCLUSION These genes were mostly first reported in Dasyatis akajei. Further studies may partly explain the molecular mechanisms by which the caudal spine of Dasyatis akajei were often used in the treatment of inflammation, parasitosis and cancer in the traditional Chinese medicine.