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OALib Journal期刊
ISSN: 2333-9721
费用：99美元
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一种新型重组免疫毒素dt390-rantes真核表达质粒的构建及功能
, PP. 1344-1347
Keywords: 免疫毒素,rantes,dt390,真核表达
Abstract:
目的构建一种新型重组免疫毒素dt390-rantes的真核表达质粒,并对这种免疫毒素的功能进行初步研究。方法通过rt-pcr的方法,从小鼠肝脏中获得rantes基因;将rantes基因插入到含有dt390片段的真核质粒srα中,构建成重组质粒;重组质粒转入大肠杆菌jm109,筛选出含有正确插入片段的克隆;限制性内切酶酶切图谱分析重组质粒,并进行dna序列分析;用脂质体介导法转染nih3t3细胞,免疫荧光检测重组质粒的表达情况;mtt法测定dt390-rantes的表达活性。结果经过酶切分析及dna测序证实,rantes基因正确插入到真核表达质粒srα中,并在nih3t3细胞中表达;mtt法证实,重组免疫毒素dt390-rantes能在体外杀伤活化的t细胞。结论成功地构建了一种新型重组免疫毒素真核表达质粒dt390-rantes-srα,该质粒可在真核细胞中瞬时表达,其转染上清对活化的t细胞具有杀伤作用。
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