查询词典 renature

We dissolve it in guanidine hydrochloride, purify it with Ni++ affinity chromatography column and renature it in vitro.

同时，使用RT-PCR只涉及引物使用与保存，不涉及VSV的使用，可以避免散毒的危险。

The enzyme reductively unfolded is almost unable to renature indicating the roles of disulfide bonds on maintaining the correct tertiary structure of the enzyme.

还原条件下变性的酶几乎不能复性，表明二硫键对于维持酶的特定空间结构是极为重要的。

Inclusion bodies are easily formed when recombinant proteins are expressed in E.coli system, and how to renature these inclusion bodies is now becoming the key problem in the genetic engineering.

E.coli作为目前应用最为广泛的原核表达系统，在异体表达蛋白质的过程中容易形成无活性的包涵体。

But 5-helix was apt to form inclusion body when expressed directly in prokaryotic cell and was difficult to renature, which causes inconvenience to future study.

但5-helix基因在原核细胞中直接表达时易形成包涵体，复性困难，给研究带来不便。

With ultrasonic wave and lysozyme, the recombination N protein is split from bacteria. We dissolve it in guanidine hydrochloride, purify it with Ni〓 affinity chromatography column and renature it in vitro. By testing the antigen of the purified N protein, it shows that its activity is very high.

建立了用超声波和溶菌酶裂解菌体，用盐酸胍溶解，Ni〓亲合层析柱纯化及体外复性等方法，经检测证明纯化的VSV重组核蛋白抗原具有较高的活性。

The DNA of Yak PrP expressed plasmid extracted from the recombinant expressed bacterium was identified by PCR amplification and two-ezyme digestion. The expressed product was obtained from the recombinant bacterium inducted under the optimized expressed condition (37℃, 1 mmol/L IPTG, 6 h), and it was determined by SDS-PAGE and Western blotting. The GST-BoPrP(23—242) fusion proteins were collected by two ways: the first is to purify and renature from the inclusion bodies; the second is to separate by SDS-PAGE.

将保存的含有牦牛PrP基因重组表达菌提取质粒DNA，进行PCR扩增和双酶切鉴定，并在优化诱导表达条件(37℃，1mmol/L IPTG，6h)下，获得的表达产物进行SDS-PAGE和Western blotting检测；将鉴定之后的重组表达菌诱导表达后，通过两种方法回收GST-BoPrP(23～242)融合蛋白：其一，是从包涵体中提取和复性GST-BoPrP(23～242)融合蛋白；其二，是通过SDS-PAGE直接分离并纯化GST-BoPrP(23～242)融合蛋白。

Two relevant sites of enzymatic digestion were added to the mTNF-α by PCR. The mTNF-α was linked to the 3'end of m/〓 in pGEX4T-1 vector. The prokaryotic expression vector pGEX4T-1m/〓-mTNF-α was constructed successfully. After induction and expression by IPTG, the expression of two kinds of fusion protein is 15% and 12% of total bacteria proteins respectively. The anti-HCC bifunctional antibodies m/〓-mTNF-α were identified by electrophoresis after the inclusion bodies were purified, denature, renature, re-purified, digested by thrombin and further purified.

采用PCR的方法在mTNF-α的两端加上所需要的酶切位点，将之连接在m／〓的3'端，构建原核表达载体pGEX4T-1 m／〓-mTNF-α，通过IPTG的诱导表达之后，两种融合蛋白的表达量分别占细菌总蛋白的15％、12％，表达产物经包涵体的纯化→变性→复性→纯化→凝血酶酶切→进一步纯化后，可以得到纯度为电泳纯的m／〓-mTNF-α抗肝癌双功能抗体。

Results The purity and following treatment of inclusion bodies of CPB were influenced by the culturing temperature after the inducement with IPTG. The solubility of inclusion bodies of rCPB in 10mol/L urea was 2~3 times higher than in 8 mol/L urea, and the renature solution with 0.75%β-ME in 10mol/L urea could increase the solubility of inclusion bodies of rCPB, comparing with that in 10mol/L urea simply.

结果诱导后生长温度不同对rCPB包涵体的纯度及后期处理均有影响；rCPB包涵体在10 mol/L尿素溶液中的溶解度比在8 mol/L 尿素溶液中提高2~3倍；添加0.75%β-巯基乙醇能显著改善rCPB包涵体的溶解效果。

METHODS:The gene of human ET1 was synthesized according to the preferential codons of E. coli, cloned into the EcoRI and SalI sites of vector pThioHisA. The recombinant plasmid pThioHisA-ET1 was constructed , sequenced and transformed into E.coli TOP10. Induced and expressed fusion protein were identified and analysed by 12% SDS-PAGE and densitometry analyses. After the elution, denature and renature, the fusion protein Thioredoxin-ET1 was obtained by ProBondTM chromatogragraphy. The purity of Thioredoxin-ET1 was detected by HPLC. Inoculate Thioredoxin-ET1 once per mouse every 2 weeks in 25, 50 and 100μg separately on 3 groups for 4 times. 10 days after last inoculation, we obtained venipuncture blood.

根据人ET1的多肽序列合成ET1基因，将其插入到pThioHisA的EcoRI和 SalI位点，重组质粒pThioHisA-ET1进行酶切鉴定及序列测定验证后转化TOP10，IPTG诱导的重组菌经SDS-PAGE检测融合蛋白Thioredoxin-ET1的表达量；表达的融合蛋白用ProBond亲合层析纯化并经HPLC鉴测其纯度；每只小鼠按25、50、100ug/次剂量的Thioredoxin-ET1每两周免疫一次，共4次，最后一次免疫10d后制备抗血清，经Western blot和ELISA检测证明Thioredoxin-ET1融合蛋白具有ET1免疫反应原性。

The sensitivity of SMMC-7721 cells to mTNF-αin vitro is not parallel with that in vivo.Key Words: hepatocellular carcinoma; genetic engineering; variable gene; clone; single chain Fv; expression; purify; denature; renature; bifunctional antibody; targeting therapy

结合MTT实验结果，我们推测：在荷肝癌（SMMC-7721）裸鼠体内，肿瘤细胞对mTNF-α的敏感性与其在体外的敏感性是不平行的，mTNF-α有可能通过阻断肿瘤的血液供应，从而杀伤肝癌细胞。

With Death guitarist Schuldiner adopting vocal duties, the band made a major impact on the scene.

随着死亡的吉他手Schuldiner接受主唱的职务，乐队在现实中树立了重要的影响。

But he could still end up breakfasting on Swiss-government issue muesli because all six are accused of nicking around 45 million pounds they should have paid to FIFA.

不过他最后仍有可能沦为瑞士政府&议事餐桌&上的一道早餐，因为这所有六个人都被指控把本应支付给国际足联的大约4500万英镑骗了个精光。