摘要
利用固相亚磷酸三酯法化学合成了人胰岛素样生长因子Ⅰ（ＩＧＦⅠ）结构基因的４条寡核苷酸片段，然后通过１、２和３、４片段末端互补配对和Ｋｌｅｎｏｗ酶促补平成为完整的ＩＧＦⅠ双链ＤＮＡ片段，通过适当的酶切后，克隆于ｐＵＣ１９中，构建了ｐＵＣＩＧＦ克隆载体。经双脱氧末端终止法测序证明，ＤＮＡ序列与我们所设计的ＩＧＦ序列完全一致。
Four oligonucleotides were synthesized according to human insulin like growth factor Ⅰ DNA sequence by means of the solid phase phosphotriester method. Two pairs of oligos 1 and 2, 3 and 4 had respectively been annealed by their 15 base complementarity, and then polymerized into two intact double stranded gene. The two genes were then inserted into a plasmid vector pUC19 after appropriate cleavage with restriction endonuclease. The resultant recombinant plasmid pUCIGF was sequenced by dideoxy chain termination method,confirming that the sequence was identical with which we anticipated.
出处
《军事医学科学院院刊》
CSCD
北大核心
1997年第1期20-23,共4页
Bulletin of the Academy of Military Medical Sciences
关键词
胰岛素样
生长因子I
基因
重组
DNA
克隆
insulin like growth factor Ⅰ
gene
DNA, recombinant
DNA sequencing
