再纯化

Our laboratory also found a gene like Drosophila intersex in the testis of Cyprinus carpio. According to the significant function played by Drosophila ix in sex-determination, we are interested in the function of this gene in carp. To study the functions of cIX further, I cloned the full-length cDNA of cIX. The recombinant protein is produced and purified to serve as the antigen to induce antibody.

本实验室同样也在鲤鱼的睪丸中发现了类似Drosophila intersex的基因，鉴於intersex在果蝇中所扮演性别决定的重要功能，对於此基因在鲤鱼中所担任的功能而感到兴趣，为了进一步探讨cIX的功能，本论文完成cIX全长cDNA，设计并建构cIX的重组DNA，再利用纯化之重组蛋白诱导抗体。

If you thought they didn't make albums like Neil Young's "Everybody Knows This is Nowhere" anymore, cue up "Ghosts of the Great Highway," and marvel over the fact that Kozelek and company are able to cross-pollinate folk, country, punk, and psychedelic influences without sounding the least bit retro, stealing the purifying flame of Crazy Horse meltdowns like "Cortez the Killer" while sounding like no one but themselves.

你认为，他们不象尼尔杨一样使集子"，每人知道这无处"再，向上的暗示是"大的公路的鬼线"，并且越过那些事实的奇迹那Kozelek 和公司能够穿过受粉人们，国家，朽木和引起幻觉影响没有发出那些重新流行一点儿最小，在只听上去是他们自己时象"科尔特斯消除剂"一样偷使燃烧纯化不含的发疯的马熔化。

Crystallized sulfuric salt of SAM was obtained by centrifuging the precipitant at 3000 rpm after adding six times the volume of absolute methanol to the decolorant liquid with a final step of freezing evaporation.

确定了分离纯化SAM的最优条件是：在pH值5.0时，含SAM浓度为3.0 g/L的上柱液，以6 L/h的流速，过装填D152树脂的离子交换柱(φ50 mm×1500 mm)后以去离子水和醋酸洗脱除杂，再用9 L到15 L浓度为50 mmol/L的硫酸以6 L/h的流速洗脱，可以分离得到纯度较高的SAM粗品。

The molecular weights of the 11 major bands were 82, 79, 60, 51, 46, 38, 32, 30, 28, 24 and 18kD, respectively. The results of Western blotting with the positive serum of anaphylactic patients showed that the 1st-4th instar larvae all had the specific allergens whose molecular weights were 82 and 79kD, respectively; only the 5th instar larvae all had the specific allergen of 30kD. We purified the protein of 30kD by ion exchange chromatography and gel isolation and identified it as ectoblast protein by MALDI-TOF-MS.

选用家蚕过敏患者阳性血清进行免疫印迹，1~4龄家蚕均显示出82和79kD的特异性变应原；但只有5龄家蚕的30kD蛋白为特异性变应原，通过离子交换层析和经切胶纯化出30kD蛋白，再经MALDI-TOF-MS鉴定该蛋白为外膜蛋白。

In some cases, its epimer 4 was also obtained. 22 compounds were synthesized, among them 18 were new.3. Using the intramolecular Pictet-Spengler reaction to form the pentacyclic skeleton of Et-743, the byproducts 25 were obtained under various conditions. The structures of them were determined to be 1, 2-dihydro-isoquinoline compounds, and the mechanism of the reaction was proposed.B. The synthesis of the analogs of Et-743Because of the failure of removing the protective groups in compounds 14, the synthetic strategy was adjusted.

对于苄基保护的羟基化合物13a，顺利得到化合物30，然后与不同的羧酸反应得到18个酯化合物33-50；对于苄氧羰基保护的氨基化合物13b，由于脱除保护基的化合物31在空气中不稳定，所以我们采取不经分离纯化，直接与羧酸反应得到酰胺化合物53-62，然后再分别对其C-21位的羰基进行还原氰基化，得到10个酰胺目标化合物63-72，另外，我们还对羟基转化为氨基的反应进行了探索。3。

Methods Level of delayed-type hypersensitivity, IL-2 of immunized BALB/c mice were determined by footpad swelling,MTT and transfered protective rate of T cells purified by neloy wool were studied with 500LD50 of the bacteria.

方法迟发型变态反应、IL-2分别采用足垫肿胀、MTT法；利用尼龙毛柱纯化免疫的BALB／c小鼠的T淋巴细胞通过尾静脉转移到非免疫小鼠，再用500LD50的STM攻击。

The protein of 14.3kDa was then analyzed by gel filtration in ion exchange chromatography, and the MALDI-TOF/TOF-MS search on the Internet. The results indicate that the protein was hemocyanin subunit.

然后用Sephadex G75凝胶过滤层析和DEAE-52离子交换层析纯化出该过敏原，再经MALDI-TOF/TOF-MS检测，利用Blastz在线分析。

Methods The total RNA was extracted from the brain of human embryo. The segment of open read frame of DOC-1 was amplified by RT-PCR and was further cloned into the vector, pMD18-T, and the prokaryotic expression vector, pGEX-4T-1, by turns, to produce the new construct pGEX-4T-1-DOC-1. After restriction enzymes digestion analysis and sequenced, pGEX-4T-1-DOC-1 was transformed into E.coli BL21（DE3）. GST-p12 recombinant protein was expressed under IPTG induction and purified by affinity column.

从人胎脑组织中提取总RNA，经逆转录-聚合酶链式反应扩增DOC-1的CDS序列，再通过基因重组技术将该基因片段依次克隆到pMD18-T和pGEX-4T-1载体中，构建融合表达载体pGEX-4T-1-DOC-1，经酶切、测序鉴定后，用该重组质粒转化E.coliBL21，用IPTG诱导表达，Glutathione Sepharose 4B柱亲和层析纯化重组蛋白。

Total RNA was isolated from cultured human pigmental cells and mRNA was reversly transcribed into cDNA. Then PCR was used to amplify the TRP -2 coding region. The PCR product was cloned into pUC19 plasmid and sequenced, then subcloned into vector pGEX-4T-1. The TRP-2 protein was expressed hi E coli of DH5 a as fusion protein with glutathioneS-transferase induced by IPTG. The fusion protein was purified by glutathione resin column.

我们通过RT—PCR的方法扩增TRP—2编码基因，克隆至pUC19载体并测序，再亚克隆至GST融合表达载体pGEX—4T—1，转化大肠杆菌，IPTG诱导表达了GST/TRP—2融合蛋白，并分析鉴定表达蛋白的可溶性，最后第四军医大学硕士学位论文一用谷氨酚胺树脂柱纯化表达的蛋白。

A complex sample containing BMP and sodium fluoride was implanted under the abdominal skin of young male Wistar rats.

在纯化的BMP中加入氟化物再植入大鼠皮下，通过比较植入物在不同植入时间的组织学变化，碱性磷酸酶、酸性磷酸酶活性及钙含量来反映BMP的生物活性。

As she looked at Warrington's manly face, and dark, melancholy eyes, she had settled in her mind that he must have been the victim of an unhappy attachment.

每逢看到沃林顿那刚毅的脸，那乌黑、忧郁的眼睛，她便会相信，他一定作过不幸的爱情的受害者。

Maybe they'll disappear into a pothole.

也许他们将在壶穴里消失