Cloning, expression, purification, and characterization of LC-1 ScFv with GFP tag

INTRODUCTION Because of its low molecular weight and ability to fluoresce independently (George, 1997), the new molecular tag, green fluorescent protein (GFP), has become more and more popular after Prasher et al.(1992) cloned its cDNA in 1992. There are many reports describing the co-expression of GFP and a specific antibody or cytokine gene, with the fusion protein expressing the fluorescent activity and bio-logical activity of the complement protein (Haraguchi et al., 1999; Mclean…     

对虾白斑综合症病毒囊膜蛋白Vp28基因的克隆、表达及蛋白纯化

以对虾白斑病毒DNA为模板扩增出Vp28基因,经限制性内切酶（SmaI,Not I）酶切后按正确的读码框顺序插入到pGEX-4T-2表达载体上,重组质粒转化大肠杆菌,经菌落PCR和质粒双酶切鉴定、序列测定确认,证实成功地构建了Vp28基因原核表达载体．转化菌经IPTG诱导表达,SDS-PAGE显示有与预期大小约52kD相吻合的融合蛋白带,18℃诱导24h后获得了可溶性表达的目的蛋白,采用Glutathione Sepharose 4B亲和层析对重组蛋白进行纯化,获得了纯度很高的目的蛋白．     

论高校网络思想道德环境的净化和建设

上网是大学生学习生活的重要内容,大量的良莠混杂的网络信息,对大学生的道德观念及价值判断造成了深刻影响,其消极方面集中表现为道德主体性的失落、道德失范和道德心理失衡。加强高校网络思想道德环境的净化和建设工作刻不容缓,这就要求净化校园的网络环境,保证网络信息健康文明的内容格调;教育学生自觉遵守网络法律规范,做文明网民‘鼓励和支持学校主管部门及“两课”教师创建专门网站,开辟高校网络思想教育新阵地。     

A case of IgA multiple myeloma: Nutritional perspective in diagnostic testing

A 64-year-old security guard and longstanding known hypertensive presented with hypertensive heart disease (HHD), weight loss, an enlarged prostate, and a spontaneously fractured rib. Malignancy of the prostate with possible metastases to the ribs was strongly suspected. Biochemical profiling revealed a paraprotein. Other biochemical and hematological correlates that were on hand before serum protein electrophoresis were rather atypical. Paraprotein studies by immunofixation revealed IgA myelomatosis. Unlike previous reports from Caucasians there was normocalcaemia, normal protein level, microcytic hypochromia, low MCHC, cholesterol level at the lower limit of the reference range and normal urea level (in the face of markedly raised creatinine level). Nutritional modulation of the classical laboratory features of this disease may account for the fairly atypical laboratory picture. The need to appreciate the influence of nutritional status on the laboratory (especially biochemical) features of a disease and thus interpretation of diagnostic tests appears of exceptional current importance, given the widening gap in socioeconomic status and the level of poverty between the resource poor and developed countries from which the classical, clinical and laboratory features of most diseases were first described. This case report reminds of the need not only to recognize theoretically the impact of nutritional status on the laboratory characteristics of a disease but of the practical application of the nutritional perspective in the interpretation of diagnostic investigations, especially in nutritionally disadvantaged communities.     

拟南芥热激因子AtHsfAla在PET原核表达系统中的表达及纯化

以pErl30a为载体构建的表达拟南芥热激因子AtHsfAla的大肠杆菌EcoliM15（pET30a／His6一AtHsfAla）为实验材料,以IPTG诱导6XHis融合蛋白的表达并经过Ni2＋柱分离纯化AtHsfAla,再通过SDS—PAGE鉴定表达蛋白和纯化蛋白．结果显示,AtHsfAla蛋白在在PET原核表达系统中能够有效表达,并能通过亲和层析获得纯化的AtHsfAla蛋白．研究结果为揭示拟南芥热激因子AtHsfAla的作用机理奠定基础．     

枇杷核中苦杏仁苷的分离纯化

以莆田"解放钟"枇杷核为原料,采用水提工艺,筛选出一种吸附性能较好的树脂,并对其进行了苦杏仁苷的分离纯化研究。结果表明采用大孔吸附树脂HZ801进行苦杏仁苷的富集纯化较佳,并确定优化枇杷核中苦杏仁苷大孔吸附树脂HZ801富集纯化工艺。     

桑树DNJ提取、纯化和检测的研究进展

1-脱氧野尻霉素(1-deoxynojirimycin,DNJ)是存在于桑树中的一种多羟基哌啶生物碱,因其通过抑制α-葡萄糖苷酶活性,进而具有降血糖、抗病毒、抗肿瘤等生理活性和药理活性.DNJ广泛存在于桑树的各种组织中,不同桑树类型和品种中DNJ含量不同,采取的每一种提取、测定方法均具有选择性和特异性,只适用于某种类型或某种桑树品种样品的纯化和检测.同时,DNJ在桑树体内的含量较低,而且桑树体内其他次生代谢物质的干扰增加了DNJ提取和测定的难度.因此,目前文献中报道的DNJ提取、分离纯化和检测方法较多.针对已报道的DNJ的提取、分离纯化和检测方法,综述了近年来DNJ研究方面的最新进展,为更好利用和开发DNJ制品,为DNJ的工业化生产提供参考和借鉴.     

博落回总生物碱的分离与纯化

本研究以博落回总生物碱含量及回收率等为考察指标,研究大孔吸附树脂分离纯化博落回总生物碱工艺。结果表明:AB-8型大孔吸附树脂对博落回总生物碱静态饱和吸附量为104.65mg/g(干树脂),洗脱率95.9%,动态饱和吸附量为96.5mg/g(干树脂),总生物碱回收率在91.24%、纯度在90%以上,是实验树脂中分离纯化博落回总生物碱的最佳大孔吸附树脂。分离纯化博落回总生物碱最佳工艺条件为:AB-8型大孔吸附树脂,洗脱剂为90%乙醇,洗脱剂用量为2～3倍树脂体积,上柱总生物碱量与树脂比为1:10.4,上柱液总生物碱浓度为21.57mg/ml,流速2～3ml/min,上柱液pH值7～8。     

耐力运动对高脂膳食诱导的脂肪肝小鼠UCP2与Mn-SOD表达及与脂肪肝关系的研究

目的:研究耐力运动对高脂膳食诱导的C57BL/6J脂肪肝小鼠肝脏UCP2及Mn-SOD表达与脂肪肝的关系.方法:以高脂膳食诱导的脂肪肝小鼠为模型,分析耐力运动20周后肝脏UCP2及Mn-SOD表达变化与肝脏脂肪变的关系.结果:肝脏UCP2与Mn-SOD在高脂膳食组(HS组)均显著升高;恢复正常膳食结合耐力运动20周组(RE组),Mn-SOD表达进一步升高,而UCP2表达显著降低.HS组形成明显的脂肪肝,恢复正常膳食加20周不运动组与恢复正常膳食结合20周耐力运动组脂肪肝得到明显改善,但RE组改善更为明显.结论:耐力运动可能以上调Mn-SOD表达而下调UCP2表达,达到既清除活性氧又减少肝脏线粒体ATP合成降低目的,从而保护肝脏免受过多活性氧和ATP耗尽的损伤.     

Considerations for protein intake in managing weight loss in athletes

Abstract

A large body of evidence now shows that higher protein intakes (2–3 times the protein Recommended Dietary Allowance (RDA) of 0.8 g/kg/d) during periods of energy restriction can enhance fat-free mass (FFM) preservation, particularly when combined with exercise. The mechanisms underpinning the FFM-sparing effect of higher protein diets remain to be fully elucidated but may relate to the maintenance of the anabolic sensitivity of skeletal muscle to protein ingestion. From a practical point of view, athletes aiming to reduce fat mass and preserve FFM should be advised to consume protein intakes in the range of ～1.8–2.7 g kg^?1 d^?1 (or ～2.3–3.1 g kg^?1 FFM) in combination with a moderate energy deficit (?500 kcal) and the performance of some form of resistance exercise. The target level of protein intake within this recommended range requires consideration of a number of case-specific factors including the athlete's body composition, habitual protein intake and broader nutrition goals. Athletes should focus on consuming high-quality protein sources, aiming to consume protein feedings evenly spaced throughout the day. Post-exercise consumption of 0.25–0.3 g protein meal^?1 from protein sources with high leucine content and rapid digestion kinetics (i.e. whey protein) is recommended to optimise exercise-induced muscle protein synthesis. When protein is consumed as part of a mixed macronutrient meal and/or before bed slightly higher protein doses may be optimal.