大肠杆菌asd基因PCR引物设计及其长模板PCR扩增

The Primers Design of the Asd Gene of Escherichia coli and its Amplification with Long Template PCR System

目的获得大肠杆菌全长ａｓｄ基因。方法采用计算机引物设计软件，长模板ＰＣＲ扩增法及限制性内切酶酶切分析。结果设计一对Ｅ．ｃｏｌｉａｓｄ基因ＰＣＲ引物；经长模板ＰＣＲ扩增获得１５１０ｂｐＰＣＲ扩增片段；经限制性内切酶酶切分析，证明该片段与电脑查询的Ｅ．ｃｏｌｉａｓｄ基因酶切谱相同。结论以本文设计的引物用长模板ＰＣＲ扩增法得到的片段初步确认为Ｅ．ｃｏｌｉａｓｄ基因。

Objective\ To obtain the asd gene of E.coli by means of PCR.\ Methods\ DNA/RNA primer analysis software was used,the fragment of asd gene was amplified with long template PCR system and digested with restrictive enzymes. \ Results\ A pair of 21mer oligonucleotides were designed.\ The 1510 bp fragment of asd gene of E.coli was amplified by using these primers and identified with restrictive enzymes,the results showed that the DNA fragments of asd gene were equal to those the “Gene Bank” shows.\ Conclusion\ The asd gene of E.coli has been obtained.\ The long template PCR system is suitable for amplification of long fragments.