人β2微球蛋白成熟肽基因克隆及其原核表达载体的构建

Gene cloning and construction of prokaryotic expression vector of Human β2-microglobulin chain mature peptide gene

根据GenBank基因库中人β2微球蛋白(β2m)成熟肽基因序列设计2对引物,使用RT-PCR法从健康人血液中扩增人β2m成熟肽基因,扩增产物进行T-A克隆和测序.结果表明,获得的人β2m成熟肽基因为297 bp,与模板序列的同源性为100%.利用基因重组技术,将β2m成熟肽基因亚克隆入pET-28 a(+)载体中,经PCR、酶切和测序鉴定,证实所获重组表达质粒pET-28/Humβ2m中含有目的片段,且连接、构建正确,表明成功构建了重组表达质粒pET-28/Humβ2m.

Two pairs of primers were designed according to the reference Human β2m mature peptide genes from GenBank.The Human β2m mature peptide gene was amplified from the blood of healthy people by using RT-PCR.PCR product was cloned into the T easy vector and sequenced.The sequencing result showed that the target gene was 297 bp and the homology between the Human β2m mature peptide gene and the template is 100%.The Human β2m mature peptide gene was subcloned into pET-28a（＋） vector.The recombinant plasmid pET-28/ Hum β2m was identified by PCR,DNA restriction and sequencing.The result showed that the recombinant plasmid pET-28/ Hum β2m was constructed correctly and successfully.