大肠杆菌N—乙酰神经氨酸裂合酶基因的组成型高效表达

ConstitutiveHigh-Expression of the N-Acetylneuraminate Lyase Gene of Escherichia coli

摘要: 应用ＰＣＲ技术从Ｅ．ｃｏｌｉＪＭ１０５中扩增出长约８９０ｂｐ的Ｎ－乙酰酶基因，将其插入高表达载体ｐＥＴ２８ａ的ＮｃｏＩ／ＢａｍＨＩ痊点，转化Ｅ．ｃｏｌｉＢＬ２１（ＤＥ３），构建高产Ｎ－乙酰神经氨酸裂合酶基因工程菌，其表达量占菌体总可溶性蛋白的７６．８％，活力比宿主菌均有不同程度的提高，其中１０７号比宿主菌高１４倍。

Abstract: bp DNA fragment encoded N acetylneuraminate lyase was amplified from E.coli JM 105 by PCR. This fragment was inserted into NcoI/BamHI sites of pET28a. The recombinant plasmids were transformed into host strain E.coli BL 21(DE3). High level expre