人β干扰素基因的改构及其融合表达和纯化

Expression and purification of human interferon-β after gene modification

目的 :通过基因改构来提高人 β干扰素基因原核表达产物的稳定性和比活性。 方法 :用RT_PCR法从人外周血淋巴细胞中获得人 β干扰素基因。定点突变后 ,克隆至表达载体pGEX_2T。IPTG诱导该基因的表达 ,亲和层析和原位裂解法纯化表达产物。结果 :获得大量纯化产物 ,且稳定性和活性获得极大提高。结论 :基因改构可以提高人

Objective:To improve the stability and the activity of human interferon_β through gene modification.Methods:Human interferon_β gene was cloned from a donor′s peripheral blood lymphocytes via extraction of total RNA, reverse transcription and nest_PCR. After site_directed mutagenesis, the gene was cloned into expression vector pGEX_2T. The modified gene was expressed by IPTG induction and purified by affinity chromatography and cleavage in place.Results:Large amount of purified gene product was obtained and its stability and activity were improved tremendously.Conclusion:The stability and activity of procaryotically expressed human interferon_β can be improved through gene modification.