摘要
目的 为进一步研究人白细胞介素 (hIL - 18)的生物学特性打下基础 ,进行hIL - 18的基因克隆及原核表达。方法 采用反转录PCR(RT -PCR)法从正常人外周血白细胞中克隆hIL - 18基因 ,并重组于PBV2 2 0表达载体上 ,经酶切初步鉴定后进行DNA序列分析 ,继而观察重组hIL - 18基因在大肠杆菌中不同诱导时间表达量的差异。结果 所克隆的基因与预期结果一致 ,并在 42℃诱导 5h时表达量最高。结论 成功地克隆了hIL - 18基因 。
Objective In order to study hIL-18 biological characteristics in a more advanced way,we proceed this study.Mehods The hIL-18 gene by RT-PCR from human peripheral white blood cells was amplified and recombined into a thermo-sensitive expression vector PBV220,then the competent cells of E.coli DH 5α was introduced into.Finally,we analyzed the different expression level of hIL-18 gene at different inducing time.Results With the identification of enzyme map and sequncing,we find our cloned gene in acconformity with the published papers about hIL-18,and the expression rate was highest when induced 5h at 42℃.Conclusion we clone hIL-18 gene successfully and obtain high rate expression.
出处
《苏州医学院学报》
2000年第9期799-801,共3页
Acta Academiae Medicinae Suzhou
