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Cloning of Human Interleukin-17 and Its Expression in E. coli
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Abstract:
Interleukin-17 (IL-17) is a newly found cytokine secreted by activated T lymphocytes. From preliminary study, IL-17 was found to play a role in the relationship between T lymphocytes and hematopoiesis, but many of its characteristics remain unknown up to now. To investigate its biological functions and related mechanisms, we cloned the complete coding region of human IL-17(hIL-17) from activated human peripheral blood mononuclear cells (PBMC) by RT-PCR and constructed an integrative vector pLCM182-hIL-17 for cloning, expressing and sequencing the hIL-17 gene. By DNA sequencing, the cloned hIL-17 is identical to the reptorted, and with temperature inducing, the hEL-17 was highly expressed in E. coli up to 30% of bacterial protein. The expressed hIL-17 protein could stimulate primary human fibrob-lasts to secrete IL-6 and GM-CSF after initial purification. This results indicate that the expressed hIL-17 has biological activity.
Keywords:
human interleukin-17 ; gene cloning ; prokaryotic expression ; interleukin-6 ; GM-CSF
Clc Number:
R392
