使复性

How to refold inclusion body proteins with many pairs of disulfide bonds into natural conformation with high efficiency is a challengeable problem at present, and it is becoming one of the industrial bottlenecks in bioengineering field.

如何对富含二硫键的包涵体蛋白进行高效的体外复性使其获得正确的空间结构是重组蛋白生产所面临的巨大挑战，现已成为制约生物工程技术产业化的瓶颈问题之一。

The inclusion bodies were solubilized in 50mM Tris-buffer at pH 12.5 and then were diluted to refold the denatured proteins. Refolding of D-hydantoinase by direct and fed-batch operations was investigated . Furthermore, the influence of the additives, CTAB and MnCl2, for soluble protein recovery was also studied.

实验中先以pH 12.5， 50mM Tris-bufferru将包涵体溶解，再藉由稀释法使其复性，并探讨直接稀释法与馈料稀释法对乙内醯尿酶复性的影响，另外探讨添加剂是否对可溶蛋白回收率有所帮助。

A new eucaryotic expression system of recombinant PoIFN-γ was also constructed so that the denaturalization and refolding process is avoided. The target gene coding PoIFN-γ2 mature protein was subcloned into expression vector, transformed into Pichia pastoris, The His〓 Mut〓 phenotype transformants were screened, fermented in flasks and induced by 1％methanol. The expressed product in culture solution and sedimentation have antiviral activity on VSV and immunological activity proved by indirected immunofluorescence antibody and Dot blotting assay.

为了构建可对表达产物修饰、加工的真核表达系统，使表达产物具有自然干扰素的活性，无需变性、复性，我们将PoIFN-γ2成熟蛋白基因连接酵母整合表达载体，电转化毕赤酵母，筛选了His〓Mut〓表型转化子，摇瓶培养，1％甲醇诱导表达，培养上清和菌体裂解上清均有抗VSV病毒活性，经间接免疫荧光抗体检测和Dot blotting鉴定，重组酵母菌诱导表达产物具有免疫活性。

The protein purified with Ni-NTA column was denatured by 8 mol/L urea and dialyzed for refolding.

Ni-NTA柱纯化后的特异性蛋白经8 mol/L脲素变性，梯度透析使蛋白质复性。

The unfolded protein correctly refolded to the native conformation through the column, and was partly purified during the elution process.

其原理是将层析技术应用于蛋白质复性和纯化，使变性蛋白质在层析柱上折叠为正确的空间构象，在洗脱的同时实现部分纯化。

The results showed that ①after the treatment with 0. 3% Triton X-100 5 min or standard binding buffer 20 min,the proteins on NCM could be detected with first specific antiserum from the titer 1:100 diluted to 1: 1000;②the blocking effect of Tween 20 was more effective than that of other blockings;③both HRP-labelled conjugate and colloid gold-labelled c...

结果表明：①经（TritonX－100）＝0.3％，5min和SBB20min的蛋白质复性处理后，可使第一免疫血清稀释度1：100改为1：1000使用；②应用（Tween20）＝0.05％作为封闭液效果最好；③金标法及酶标法都可用于Western印迹技术，酶标法主带清晰，而金标法灵敏度高。

After the sequence and open reading frame were confirmed by DNA sequencing, the GADes gene was expressed in E.coli. SDS-PAGE analysis suggested that the bacteria BL21 produce a kind of fusion protein of 92KD and the expressed GADes fusion protein shows expected immunoreactivity by Western Blot.

SDS-PAGE电泳和Western Blot分别鉴定融合蛋白大小和免疫活性后，对表达条件进行优化，对包涵体进行分离、溶解和复性处理，以提高可溶性融合蛋白的产量，使融合蛋白达总蛋白的50％，相当于1.495g/每升培养物。

According to structure analysis, the contents of α-helix and dimer structure in refolded eMDH were increased by the cooperation, and eMDH activity was enhanced.

对复性产物结构的初步分析发现，耦合作用可使eMDH分子的α-螺旋及二聚体含量明显增多，因此复性产物的活性也显著提高。

The results showed that the purified recombinant protein under denaturing conditions was refolded in Ni-NTA column by using refolding buffer.

实验结果表明，在变性条件下纯化的样品在柱上不经洗脱，直接用复性液洗涤6h，可以使EKLC在得到提纯的同时实现复性，得到了电泳纯度为96％的具有生物活性的EKLC，最高活性为712U。

The trial results by statistic analysis show that the fertilizer effect of organo-inorganic compound fertilizer is nearly same to that of inorganic compound fertilizer, the fast release of commercial organic fertilizer is also similar to that of chemical fertilizer.

通过茄子在无肥、商品有机肥、无机复混肥和有机无机复混肥4处理的一次基肥使用试验，对产量、开展度、花蕾数、结果数和单果量考察和统计分析反映出：有机无机复混肥与无机复混肥的肥效基本相同，商品有机肥似具有与化肥一样的速效性，要使有机肥发挥肥效，只有增加有机肥使用量，不能期望通过有机无机复混肥中加入少量有机肥来体现有机肥的肥效。

It has been put forward that there exists single Ball point and double Ball points on the symmetrical connecting-rod curves of equilateral mechanisms.

从鲍尔点的形成原理出发，分析对称连杆曲线上鲍尔点的产生条件，提出等边机构的对称连杆曲线上有单鲍尔点和双鲍尔点。

The factory affiliated to the Group primarily manufactures multiple-purpose pincers, baking kits, knives, scissors, kitchenware, gardening tools and beauty care kits as well as other hardware tools, the annual production value of which reaches US$ 30 million dollars.

集团所属工厂主要生产多用钳、烤具、刀具、剪刀、厨具、花园工具、美容套等五金产品，年生产总值3000万美元，产品价廉物美、选料上乘、质量保证，深受国内外客户的青睐