摘要
通过在不同温度、不同诱导时间、不同异丙基硫代 β Ｄ 半乳糖苷（ＩＰＴＧ） 诱导浓度条件下，对重组大肠杆菌ＰＰＳＬＴ ⅡｅＢ 表达的谷胱甘肽 Ｓ 转移酶（ＧＳＴ） 与类志贺氏菌毒素Ⅱ型变异体Ｂ 亚单位（ＳＬＴ ⅡｅＢ） 的融合蛋白（ＧＳＴ ＳＬＴ ⅡｅＢ） 的分析，结果表明：在所试条件中，温度是影响表达的主要因素，重组大肠杆菌（ＰＰＳＬＴ ⅡｅＢ） 在２８ ℃温度条件下，可以明显表达融合蛋白ＧＳＴ ＳＬＴ ⅡｅＢ． 在ＩＰＴＧ 为１ｍｍｏｌ·Ｌ－ １ 浓度条件下，２ ｈ 的诱导即可获得明显表达． 在所试几种ＩＰＴＧ 诱导浓度下，通过５ ｈ 的诱导，均可获得明显表达．４００ ｍＬ２ＸＹＴＡ 培养基产生的融合蛋白在１ ．８ ｍｇ 以上．
Recombined plasmid ppsltⅡeB which contained the sltⅡeB gene was constructed, and SLTⅡeB (7.568 kd) was expressed as a fusion protein linked with the enzyme glutathione S transferase (GST, 27.335 kd). The SLTⅡeB fusion protein was designated as GSTSLTⅡeB (34.885 kd). The effects of culture conditions on production of GSTSLTⅡeB were examined. Incubation time, incubation temperature, and IPTG concentrations were investigated. Culture condition that resulted in the greatest yield of GSTSLTⅡeB were incubation at 28℃. The optimal incubation time for production of GSTSLTⅡeB under the condition used in this study was 2 h after induced by 1.0 mmol·L -1 IPTG. The optimal yield of GSTSLTⅡeB could obtained after 5 h induced by different IPTG concentrations that used in this study. The data of this study also show that GSTSLTⅡeB yields was at least 1.8 mg per 400 mL 2XYTA medium.
出处
《扬州大学学报（自然科学版）》
CAS
CSCD
1999年第4期27-30,共4页
Journal of Yangzhou University：Natural Science Edition
基金
江苏省自然科学基金
