新城疫病毒HN基因片段原核表达重组质粒的构建

Construction of the Prokaryotic Expression Recombinant Plasmids Containing NDV Gene Fragments

根据GenBank中发表的新城疫病毒F48E9基因序列设计引物,采用RT-PCR技术,成功获得了该病毒HN基因中两个不交叉的基因片段,长度分别为790bp和579bp。将这两段基因分别克隆到pGEM-T vector中,经酶切鉴定和测序后克隆到原核表达载体pET-28a,PCR和酶切鉴定表明,克隆的两个新城疫病毒基因片段已经正确地插入到了原核表达载体pET-28a之中。

Using Primers which were designed according to the sequence of HN gene of NDV F48E9 strain, two cDNA fragments 579 bp and 790 bp in length,of HN gene were successfully amplified by RT-PCR, and they are 579 bp and 790 bp in length. These two amplified CDNA fragments were cloned into pGEM-T vector. The results of identification by restriction endonuclease and sequencing analysis showed that they were successfully inserted into the expression vector pET-28a.