eglinC基因的化学合成与克隆

eglinC基因的化学合成与克隆 CHEMICAL SYNTHESIS AND CLONING EGLIN C GENE 傅敏杰 1 2 3 first-author 张晓东 1 2 3 王鄂生 1 2 3 申宗侯 1 2 3 王启松 1 2 3 闵永洁 1 2 3 武汉大学生化系 武汉大学生化系 School of Life Sciences, Wuhan University School of Life Sciences, Wuhan University 复旦大学遗传所 复旦大学遗传所 武汉大学生化系复旦大学遗传所 武汉大学生化系复旦大学遗传所 采用固相亚磷酰胺法合成了ｅｇｌｉｎＣ全基因，该基因全长２２１ｂｐ，包括其结构基因，５＇－端起始密码子ＡＴＧ和３＇－端终止密码子ＴＡＧ，并在基因的５＇－和３＇－分别装上ＥｃｏＲＩ和ＢａｍＨＩ位点．共分１２个片段，采取分段合成，酶促连接成完整的ｅｇｌｉｎＣ基因．合成的基因克隆于Ｍ１３载体，经杂交、检测，筛选出含ｅｇｌｉｎＣ基因的克隆．用双脱氧链终止法测其序列，结果与设计的一致． A total gene for eglin C has been chemically synthesized by using the solid phosphoramidite method. The gene is 221 bp in length containing the structure gene encoding 70 amino acids,a start coden ATG at 5’-end and a stop coden TAG at 3’-end of the structure gene,except two restriction sites at the ends. The synthesiS of the gene involved enzymatic joining synthetic oligonucleotiede fragments to form DNA duplexes, cloning into vector M13, idnetifying by dot bloting. Finally,the synthetic coding sequence was fully verified right by dideoxynucleotide chain termination method. eglinC基因 化学合成 克隆载体 点杂交 DNA序列分析 eglin C gene chemical synthesis cloning into vector dot bloting DNA sequencing Q342.2 湖北省自然科学基金(2017CFA007) 1994-03-01 2021-04-01 3