通过T-A克隆技术,成功地构建了克隆载体pMD-18T-EGF,用EcoRⅠ和HindⅢ双酶切克隆质粒pMD-18T-EGF后,回收EGF基因,并将此基因定向克隆至相同双酶切回收后的pET32a原核表达载体中,获得重组质粒pET32a-EGF。经限制性内切酶分析和PCR鉴定,结果表明：成功地构建了原核重组质粒。%The clone vector pMD-18T-EGF was constructed successfully by T-A clone technique and was digested with EcoR Ⅰ and HindⅢ, and the prokaryotic expression vector pE332a was also cut, then the gene was cloned into the vector. Finally a recombinant plasmid named pET32a-EGF was obtained, and identified by PCR, restriction enzyme analysis and sequencing.
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