摘要
利用DNA合成技术,设计并合成人红细胞生成素(EPO)的全基因,EPO基因全长600 bp.首先合成32个小片段,经纯化后分别连接成相应的3个基因大片段,再将3个基因大片段组装成1个完整的EPO基因。用双脱氧末端终止法对基因大片段及全基因进行DNA序列分析,结果表明化学合成的基因序列与最初设计的EPO基因序列完全一致。
A 600 bp synthetic erythropoietin (EPO) gene encoding all 166 amino acids of the EPO protein and 27 amino acids of the signal peptide has been constructed. The whole gene was divided into three large fragments consisting of a total of 32 oligonucleotides. These oligonucleotides were synthesized by the solid-phase phosphor-amidite method and ligated into three large fragments. These latter three were separately cloned into vector M13mp19 and then transformed into E.coli JM 103. Positive clones were screened with 32P-labeled probes. The sequences of the fragments were confirmed by DNA sequencing, and the sequence of the whole synthetic EPO gene was confirmed by enzymatic digestion and sequencing. The results indicated that the nucl-eotide sequence of the synthetic EPO gene is identical to that of the original.
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
1992年第3期173-178,共6页
Acta Academiae Medicinae Sinicae
基金
国家"七五"重点课题
