CAJ | 학술논문

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-28a(+)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/Ham β2m.
근거GenBank기인고중인β2미구단백(β2m)성숙태기인서렬설계2대인물,사용RT-PCR법종건강인혈액중확증인β2m성숙태기인,확증산물진행T-A극륭화측서.결과표명,획득적인β2m성숙태기인위297 bp,여모판서렬적동원성위100%.이용기인중조기술,장β2m성숙태기인아극륭입pET-28a(+)재체중,경PCR、매절화측서감정,증실소획중조표체질립pET-28/Humβ2m중함유목적편단,차련접、구건정학,표명성공구건료중조표체질립pET-28/Ham β2m.
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 sequen-cing 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 re-striction and sequencing.The result showed that the recombinant plasmid pET-28/Hum β2m was con-structed correctly and successfully.