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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 ,并使获得高效表达
Objective To further study the biological characteristics of human interleukin (hIL - 18) and to clone and express hIL - 18 gene. Methods The hIL - 18 gene was cloned from normal human peripheral blood leukocytes by reverse transcription polymerase chain reaction (RT - PCR) and was then recombined on the PBV220 expression vector. The recombinant plasmid was identified by restriction enzyme digestion and DNA sequence analysis. The recombinant hIL - - 18 gene in Escherichia coli in different induction time of expression differences. Results The cloned genes were consistent with the expected results and expressed the highest at 5h after induction at 42 ℃. Conclusion The hIL - 18 gene was successfully cloned and expressed efficiently