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双语推荐：GroE

构建甘精胰岛素(ING)表达载体,提高ING在大肠杆菌中的可溶性表达。方法:在传统方法中ING的A、B链之间用C肽连接,本实验在ING的A、B链之间插入大肠杆菌硫氧还蛋白质(Trx)来替代C肽,进行ING的融合表达(ING-Trx),并进一步使用多分子伴侣GroEL、GroES、触发因子进行共表达。结果:可溶性的ING-Trx融合蛋白约占该目的蛋白质总表达量的35%;多分子伴侣共表达后可溶性ING-Trx的产量提高到ING-Trx总蛋白的78%,初步纯化后其产量为4.5 mg·L-1,是使用分子伴侣前的3倍。结论:用Trx代替C肽不影响A、B链的有效折叠,采用分子伴侣共表达可以显著提高目的蛋白的可溶性。

Objective:To construct expression vector of insulin glargine ( ING) and increase soluble expression of ING.Methods: C peptide was used to join A chain and B chain of ING in previous studies .In the present research , thioredoxin ( Trx) was inserted between A chain and B chain of ING to replace C peptide .Co-expression of molecular chaperones GroEL , GroES, trigger factor and ING was also used to promote folding of recombinant ING-Trx protein.Results:Soluble ING-Trx accounts for 35% of total expressed ING-Trx while soluble ING-Trx increased to 78%of total ING-Trx protein after co-expression.The yield was about 4.5 mg· L-1 after preliminary purification , three times as much as the production without co-expression of molecular chaperones .Conclusion:In summary , A chain and B chain can fold properly when Trx is used to replace C peptide .Co-expression of ING-Trx with molecular chaperones can greatly increase soluble expression of insulin glargine .

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目的：观察耐青霉素肺炎链球菌异常蛋白表达，探讨其蛋白组学差异与耐青霉素的关系。方法采用次抑菌浓度法将肺炎链球菌国际标准菌株诱导为耐青霉素肺炎链球菌株；双向电泳分离肺炎链球菌标准株和诱导耐药株的全菌蛋白质；Image Master 2D Platinum 5.0软件对电泳图谱进行分析，寻找表达差异蛋白点；对差异蛋白点进行质谱分析。结果成功诱导耐青霉素肺炎链球菌，并建立肺炎链球菌标准株和耐药株的全菌蛋白双向电泳图谱。标准株和耐药株分别分离出约320，350个蛋白点，发现10个差异蛋白。与标准株比较，ABC转运器、触发因子、DNA聚合酶Ⅲ、氨基酸ABC转运和SP表面蛋白A表达量上调，分子伴侣、脂蛋白、果糖二磷酸醛缩酶、α-烯醇化酶和假想蛋白表达量下调。结论耐青霉素肺炎链球菌蛋白表达异常导致的蛋白组学改变可为深入探讨其耐药机制提供新的研究方向。

Objective To analyze the proteomics of penicil in- resistant streptococcus pneumonia (PRSP). Methods The penicil in- resistant streptococcus pneumonia strain was induced by sub- MIC method. The protein profile of Streptococcus pneumoniae and induced PRSP were examined by two- dimensional electrophoresis;and the electrophoretic patterns were ana-lyzed by using Image Master 2D Platinum 5.0. The differential y expressed proteins were further identified by mass spectrometry. Results PRSP strains were induce successful y. Ten differential y expressed proteins were identified between Streptococcus pneumoniae and induced PRSP. The expression of ABC transporter, trigger factor, DNA Polymerase III, amino acid transport, pneumococcal surface protein A was increase in PRSP;while the expression of chaperonin GroES, lipoprotein, fructose bisphos-phate aldolase,α- enolase decreases, hypothetical protein SpneT_02001699 was decreased. Conclusion The findings of pro-teomics change in penicil