摘要
本文报道用化学法全合成降钙素的基因。基因全长113碱基对。采用大肠杆菌偏爱密码子。整个基因分为6个片段合成。用T^4DNA连接酶一次连接成功,序列分析证明合成的基因序列与设计的一致。
Calcitonin has been used in clinic for bone diseases etc..This paper describes a total chemically synthesis of a gene for calcitonin.The synthetic gene contains the structure gene, a EcoR Ⅰ site at 5' end, a Hind Ⅲ site and stop signal at 3 ' end.Besides these, two methionine codonswere placed at both ends of the gene, and methionine codon at 8 th position from N-terminus of human calcitonin was replaced by valine codon.Thus, after the expression and treatment with cyanogen bromide followed with ammonia an entire calcitonin carrying amide group at its C-terminus can be obtained.The total gene was assembled by a single addition of six oligodeoxy-nucleotides which was synthesized by phosphoramidite method on support. After a ligation of synthetic gene to Ml3mpl9 and transformation into E.coli, the phage bearing calcitonin gene was selected by hybridization.The synthetic gene was determined by DNA sequencing.
出处
《基础医学与临床》
CSCD
1990年第5期28-31,共4页
Basic and Clinical Medicine
关键词
降钙素
基因合成
基因克隆
calcitonin gene synthesis gene cloning
