重组人IL-6在沙门菌x4550中的表达

Expression of recombinatant human interleukin 6 in salmomella typhimurium x4550

为在鼠伤寒沙门菌ｘ４５５０中表达重组人白细胞介素６（ＩＬ６），利用定点突变、聚合酶链反应、体外重组ＤＮＡ技术构建出重组表达载体ｐＹＡ３３３３ＩＬ６；利用ＩＬ６依赖性的小鼠杂交瘤细胞株Ｂ９测定重组人ＩＬ６ＨＧＦ的活性。结果：从全菌ＳＤＳＰＡＧＥ考马斯亮蓝染色后薄层扫描分析表达产物产率为１７６％，相对分子质量为２１×１０４；测定重组人ＩＬ６ＨＧＦ活性为２×１０６Ｕ／Ｌ。提示：鼠伤寒沙门菌ｘ４５５０能够表达人ＩＬ６。

In order to express recombinant human interleukin 6 (rhIL 6) in Salmonella typhimurium x4550,a recombinant expression plasmid pYA3333 IL6 was constructed by using PCR amplification,site specific mutagenesis and DNA recombination in vitro.Hybridoma growth factor (HGF) activity of rhIL 6 was measured with an IL 6 dependent B cell hybridoma line B9.Results showed that the expressed rhIL 6 relative molecular weight 2 1×10 4 was accounted for 17.6% of the S.typhimurium proteins and the biological activity of the expressed rhIL 6 was 2×10 6 U/L bacterial medium,suggesting that salmonella typhimurium x4550 could be used to express recombinant human interleukin 6.