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To express chicken osteoprotegerin in Pichia pastoris and determine its inhibitive effects on mature osteoclastic bone resorptive function in vitro. The chOPG cDNA encoding for 382 amino acid without signal peptice was subcloned into Pichia pastoris expression vector pPICZα-A. The constructed plasmid was transformed into yeast X-33 by electroporation. The recombinant transformants were selected by Zeocin. Induced by the addition of methanol every 24 hours, the product analyzed by SDS-PAGE was sized about 43kDa and 53kDa at a yield of 200mg per litter of culture. The result of Western blotting indicated that the recombinant protein had specific antigenicity mainly owing to heterogeneous glycosylation.

为了在毕赤酵母中分泌表达鸡骨保护素（osteoprotegerin，OPG），采用DNA重组技术将chOPG成熟肽cDNA片段插入毕赤酵母表达载体pPICZα-A中，以电穿孔法转化酵母X-33，用Zeocin 平板筛选重组子，经甲醇诱导表达后， SDS-PAGE 和免疫印迹分析表达产物，由于糖基化不同，表达产物有两种，其相对分子量分别为43kDa和53kDa，表达量约为200mg/L，经Western印迹验证，有较好的抗原性。

The purified protein induced as antigen was used to immunize the rabbit, and the antiserum of WMV CP was prepared whose titer measured by ELISA was 1:6400; western blot analysis indicated that the antiserum could serologically react with CP.

以诱导的蛋白为抗原免疫家兔，制备了WMV CP抗血清，ELISA法测其效价为1/6400，Western blot分析能与CP发生血清学反应。

A complete bovine 18ku-bFGF gene was cloned by nested-PCR and subcloned into expression vector pET-28a.The recombinant plasmid pET-28a-bFGF was transformed into Escherichia coli BL21.At 25℃,recombinant protein was induced by 0.5mmol/L IPTG for 5 hours.Supernatant of cell lysate was purified using Ni-NTA method and the products was submitted to detect the bioactivity by Western-blotting,which indicated that the fusion protein contained recombinant bovine bFGF.Results showed that recombinant bovine bFGF was produced and solubly existed in the supernatant.NIH-3T3 fibroblasts were used to detect the bioactivity of the fusion protein.

采用巢式PCR方法克隆了牛18ku-bFGF基因完整的编码序列，并构建了原核表达载体pET-28a-bFGF，将其转化大肠杆菌BL21，在25℃低温条件下，用0.5mmol/L IPTG诱导表达5h，用Ni-NTA亲和纯化细胞裂解上清液，经Western-blotting检测，结果显示，在特定的诱导条件下，重组牛bFGF基因在大肠杆菌中获得了表达，并且主要以可溶性状态存在于细胞中。

Methods By the case-control study, 76 cases of Cushing ulcer after cerebral infarction and 80 cases of similar patients condition cerebral infarction without Cushing ulcer were selected. Hp-IgG was detected by Western blot in the patient's blood serum, at the same time to survey blood clotting tetrachoric level.

选择76例脑梗死并发应激性溃疡患者作为观察组，筛选病情相似的单纯脑梗死患者80例作为对照组，用Western blot法测定患者血清特异性抗体Hp-IgG，同时测量凝血四项水平。

To elucidate the possible mechanism of differentiation and/or apoptosis in NB4, K562 cells induced by tributyrin, a histone deacetylase inhibitor , the level of acetylated histone H3 was detected by Western blot, p21(superscript WAF1) expression was detected by semi-quantitative RT-PCR.

为了探讨组蛋白去乙酰化酶抑制剂三丁酸甘油酯(tributyrin， TB)诱导NB4、K562白血病细胞分化和凋亡的作用机制，利用Western blot方法及逆转录聚合酶链反应检测TB作用前后NB4、K562细胞组蛋白H3乙酰化水平以及p21(上标 WAF1)表达量的改变。

Methods The ES antigens of Trichinella spiralis muscle larvae cultured in vitro at 18 h and 30 h were analyzed by SDS-PAGE and Western blotting.

应用SDSPAGE和Western印迹对旋毛虫肌幼虫体外培养18、30h后的ES抗原中的蛋白组分进行研究。

Through SDS-PAGE and Western blot analysis, it was indicated that the expressed products were specifically detected by positive serum of mouse against Trichinella.

SDS-PAGE电泳显示表达蛋白为36ku的融合蛋白，将表达产物进行Western blot检测，可以被小鼠旋毛虫的阳性血清识别。

Methods Endogenous EB1 mRNA and protein levels in embryonic cells of trophonema from artificial abortion and spontaneous abortion were compared, using fluorescence quantitative PCR, immunohistochemistry and western blot, respectively.

采用荧光定量PCR技术检测EB1基因在人工流产绒毛核型分析正常的胚胎细胞和自然流产绒毛核型分析异常的胚胎细胞中的表达差异；免疫组织化学方法检测EB1蛋白在两者中的表达情况，并用Western blot加以验证。

To get more information of the function of human Era on cell cycle, the complete control inducible mammalian expression system was used to express human Era mutant and analyzed cell cycle of uninduced (add 1% ethanol) and induced (10μM Pon A) cell lines by flowcytometer, the primary results demonstrated human Era mutant could inhibit apoptosis induced by ethanol, and Era S36N may stimulate the expression of Bcl-2 as detected by Western Blot.

在构建稳定高表达蜕皮激素受体基因的细胞株3V3的基础上，转染突变型人Era的表达载体进行压力筛选，对所得到的单克隆细胞株进行诱导与未诱导条件下细胞周期的分析，结果表明未诱导加酒精的对照细胞可以引起细胞凋亡，而Era诱导表达后可以显著抑制凋亡细胞的数量，Western表明Era S36N可以诱导细胞Bcl-2表达，提示人Era可能与细胞凋亡相关。

The mc2155 carrying pMind-glmU-AS was induced by different concentration of tetracycline, and GlmU protein was detected in the mc2155 carrying pMind-glmU-AS by Western blotting. The results showed that tetracycline of 20ng/ml has obviously inhibited the growth rate of mc2155 carrying pMind-glmU-AS, but the amount of GlmU protein did not change compared to that of uninduced mc2155 carrying pMind-glmU-AS.

glmU反义RNA的诱导表达与GlmU蛋白的检测分别用5 ng/ml、10 ng/ml和20 ng/ml的四环素诱导携带pMind-glmU-AS表达载体的mc~2155菌株表达反义RNA，绘制细菌生长曲线，结果表明20 ng/ml浓度的四环素可抑制mc~2155/pMind-glmU-AS的生长，但Western Blotting分析结果显示经四环素诱导和未诱导的mc~2155/pMind-glmU-AS中，GlmU蛋白表达量无明显的变化。

For a big chunk of credit-card losses; the number of filings (and thus charge-off rates) would be rising again, whether

年美国个人破产法的一个改动使得破产登记急速下降，而后引起了信用卡大规模的亏损。

Eph. 4:23 And that you be renewed in the spirit of your mind

弗四23 而在你们心思的灵里得以更新