人IL-6 /sIL-6R 结合分子模型的建立及在药物筛选中的应用

目的：建立人IL-6 /sIL-6R 结合的分子模型,用于筛选IL-6 /sIL-6R的抑制剂。方法：将人IL-6基因克隆至原核表达载体pET28a（+）中表达IL-6蛋白,western blot及人IL-6检测试剂盒分析鉴定表达蛋白。同法将人sIL-6R在pET15b载体中表达,纯化并用western blot检测目的蛋白。依据ELISA原理建立IL-6 /sIL-6R 结合的分子模型,并通过改变IL-6、sIL-6R及IL-6 antibody的浓度来优化该模型,用于IL-6 /sIL-6R拮抗药物的筛选。结果：人IL-6可在载体PET28a（+）中高效表达,且经western blot鉴定正确,人IL-6检测试剂盒检测显示具有较高的免疫活性。sIL-6R在PET15b中表达,western blot鉴定正确。通过对IL-6 /sIL-6R结合的分子模型的优化,得到其最佳条件为：IL-6R 1?g/well, IL-6 500ng/well, IL-6 antibody 1?g/well。应用该模型筛选发现有些化合物可显著抑制IL-6与其受体的结合。结论：成功构建IL-6 /sIL-6R结合的分子模型,为高通量筛选IL-6拮抗剂提供平台。     

6C6免疫毒素的原核表达、纯化及其对人乳腺癌细胞的作用研究

在人乳腺癌和其他肿瘤细胞表面通常过表达一 2 8ku膜蛋白 ,可作为肿瘤导向治疗的作用靶 .利用基因工程技术 ,将 6C6单链抗体 (ScFv6C6)同缺失细胞结合区的绿脓杆菌外毒素A(PE40 )相连 ,构建成 6C6免疫毒素 (ScFv6C6-PE40 ) ,其中ScFv6C6由能特异识别 2 8ku蛋白的 6C6单克隆抗体的重链可变区和轻链可变区连接而成 . 6C6免疫毒素的原核表达水平为 3 3 % ,约 5 5mg/L菌液 .利用HisTrap(镍离子螯合 )柱层析纯化 ,得到纯度为 3 3 2 %的 6C6免疫毒素 ,其可识别MDA 2 3 1人乳腺癌细胞表面上的肿瘤相关抗原 ,并可杀伤这些细胞 ,半致死剂量为92ng/mL .     

抗果蝇Dxl6蛋白抗体的制备及特异性分析

为了研究果蝇中SR蛋白家族新成员Dxl6的功能,通过RT-PCR得到Dxl6的全长cDNA,并根据Dxl6基因产物的功能区,分别将Dxl6中间区域（Dxl6 middle part, Dxl6MP）、Dxl6 C末端RS结构域（Dxl6 RS domain, Dxl6RSD）序列亚克隆至pGEX-4T-1(His)6C 及pET32a 表达载体中,表达和纯化获得融合蛋白。用纯化的融合蛋白GST-Dxl6RSD-His和GST-Dxl6MP-His免疫家兔,分别得到抗Dxl6RSD和抗Dxl6MP两种抗体。WESTERN BLOT结果显示两种抗体能特异地识别在原核表达系统内表达的抗原,抗Dxl6RSD的抗体对果蝇组织中的Dxl6具有较高的特异性。Abstract: In order to study the function of Dxl6 which is a novel member of SR protein family, its cDNA was cloned by RT-PCR, and the sequences of its RS domain and its middle part were subcloned into two fusion express vectors, pGEX-4T-1His（6）C and pET32a. After expressing in E.coli BL21, the truncated proteins of Dxl6 RS domain part and Dxl6 middle part in pGEX-4T-1His（6）C were purified and used to immunize rabbits. Purified antibodies against the RS domain and Dxl6 middle part were obtained by affinity chromatography with the expressed products of Dxl6 RS domain part and Dxl6 middle part in pET32a, respectively. The result shows the antibody against Dxl6 RS domain has a good specificity to Dxl6 in Drosophila larvae by Western blot analysis.     

6C6鼠-人嵌合抗体的纯化及鉴定

采用基因工程技术 ,将小鼠 6C6单克隆抗体可变区基因与人抗体恒定区基因连接 ,构建了鼠 人 6C6嵌合抗体基因 ,并在CHO细胞中高效表达 .利用ProteinA亲和层析柱从细胞培养上清中分离纯化 6C6嵌合抗体 ,得到电泳纯度大于 98%的 6C6嵌合抗体 ,其重链 (5 5kD)和轻链 (2 4kD)符合IgG相对分子质量的理论值 .Western印迹、细胞免疫荧光和免疫组织化学实验结果均呈阳性 .表明6C6嵌合抗体可识别人乳腺癌细胞表面上的肿瘤相关抗原 ,保持了 6C6单克隆抗体的特性 ,为后续的研究工作奠定了基础     

佩妃延腹小蜂属榕小蜂CYP6家族进化分析

采用PCR方法获得了佩妃延腹小蜂属Philotrypesis榕小蜂9条CYP6基因片段：CYP6CK1, CYP6AQ2, CYP6AQ3, CYP6AS20, CYP6AS21, CYP6AS22, CYP6AS23, CYP6AS24和 CYP6AS25。这些基因序列都包含编码蛋白疏水螺旋区Ⅰ和血红素结合域的区域。使用MEGA 4.0构建基因树,用PAML 4.0检测选择压力。结果表明：榕小蜂的CYP6AS, CYP6AQ, CYP6CK基因亚家族的扩张可能是在自然选择下通过基因复制方式完成的。基于最大似然法的位点特异性模型对蛋白编码序列选择压力检测发现：榕小蜂CYP6AS亚家族的基因接受净化选择,在氨基酸序列上表现出较高的保守性。     

Dual-Targeting of Arabidopsis 6-Phosphogluconolactonase 3 （PGL3）to Chloroplasts and Peroxisomes Involves Interaction with Trx m2 in the Cytosol

The oxidative pentose-phosphate pathway （OPPP） represents a central branch of cellular metabolism emanating from glucose-6-phosphate （G6P） to provide reductive power （NADPH） and sugar phosphates for anabolic biosyntheses. In plant cells, the oxidative OPPP branch is found in the cytosol and in plastids, consisting of glucose-6-phosphate dehydrogenase （G6PD）, 6-phosphogluconolactonase （PGL）, and 6-phosphogluconate dehydrogenase （6PGD）. These enzymes are encoded by small gene families in the nuclear genome, which, in Arabidopsis, comprise six G6PD, five 6-PGL, and three 6-PGD isoforms （Kruger and von Schaewen, 2003）. Specific targeting motifs at the C-terminus of 6-PGL and 6-PGD isoforms suggested that the OPPP may also occur in peroxisomes （Reumann et al., 2004）.     

小麦——黑麦易位系的研究

为了创制在小麦生产上有经济价值和在小麦育种中有利用价值的小麦—黑麦易位系,用小麦—黑麦代换系5R/5A与6R/6A杂交,在杂种后代中选择带有某些黑麦性状的普通小麦类型植株,并按性状继代选择直至稳定为止,至F6代共选出9个品系即6-26、06-60-11、06-6-24、06-6-15、06-6-35、6-34、6-28、06-6-17、06-6-14。本文对这9个品系及其亲本进行了田间观察和遗传分析,结果表明：它们田间表现生长整齐一致、遗传稳定、育性正常,具有大穗、多小穗、多分蘖、抗病、抗旱等优良性状;选择分布在黑麦5R和6R染色体上的微卫星引物共20对,结果表明引物SCM268在6-26、06-60-11、06-6-15、06-6-35、6-34、6-28、06-6-17、06-6-14这8个品系中扩增出黑麦特异产物,初步确定这8个品系是易位系,是有经济价值和利用价值的遗传材料。     

hIL-6·hIL-6Rα·gp130三元复合物的结构预测

通过Delphi方法分析人白介素6（human interleukin-6,hIL-6）／人白介素6受体α亚基（human interleukin-6 receptor α subunit,hIL-6Rα）复合物、gp130（β subunit）的空间构象的表观静电分布,利用分子对接方法研究gp130与hIL-6/hIL-6Rα复合物作用形成三元复合物的空间构象,经过分子力学优化、分子动力学常温动态模拟借助分子间相互作用（范德华力、氢键、盐键等）、反应自由能理论探讨hIL-6·hIL-6Rα·gp130复合物稳定构象结合部位的结构域.分析结果表明,gp130蛋白表面富集较强的负电势,复合物hIL-6/hIL-6R一侧表面富集较强的正电势,gp130借助蛋白表面的静电作用结合hIL-6/hIL-6R复合物,介导hIL-6信号;hIL-6中的helix-C、loop-BC、loop-CD参与同gp130中的loopEF、linker、loopA′B′、loopB′C′、loopD′E′作用,hIL-6R中的loopA′B′、β-strand E′参与同gp130中的loopA′B′、β-strand E′作用.     

克隆、表达及纯化核糖体蛋白S6用于体外激酶活性检测

目的:构建40S核糖体蛋白S6的原核表达载体,表达并纯化S6蛋白,将其作为底物用于S6激酶(S6K)的体外活性测定。方法:采用RT-PCR方法从人胚肾细胞HEK293中获取S6 cDNA,将扩增产物克隆至大肠杆菌表达载体中,进行酶切及测序鉴定;IPTG诱导GST-S6融合蛋白在大肠杆菌中表达,用谷胱甘肽亲和层析纯化GST-S6,免疫沉淀法检测该蛋白是否可作为底物用于S6K的体外激酶活性测定。结果:酶切及测序鉴定表明构建了S6原核表达载体,并表达及纯化出GST-S6融合蛋白,相对分子质量为55×103。该蛋白可用于S6K的体外激酶活性测定,特异性强。结论:S6蛋白的克隆、表达与纯化成功,可用于S6K的体外激酶活性测定,为研究S6K的功能奠定了基础。     

小麦—簇毛麦染色体代换系、易位系特异蛋白组分的双向电泳比较分析

利用双向电泳技术,对栽培小麦(AABBDD)、染色体代换系(6V/6A)、易位系(6VS/6AL)、(6VS/6DL)和簇毛麦(VV)的叶片全蛋白进行了比较研究。在栽培小麦、代换系和两个易位系中检测到超过350个蛋白组分,它们的分子量范围是10~110 KD,等电点在4.5～8.6之间。栽培小麦、6V/6A、6VS/6AL、与6VS/6DL之间的双向电泳谱型极为相似,但与簇毛麦不同。在代换系、两个易位系和簇毛麦中检测到了特异蛋白组分16 KD/pI5.0,而在栽培小麦中未检测到该组分,这些结果表明16 KD/pI5.0蛋白可能定位于簇毛麦V染色体短臂上。     

韩国华津浦湖的鱼类区系

There are sevenlagoons onthe east coast of Korea,including Kyeongpo Lake,Hyang Lake,Mae Lake,Chung-cho Lake,Yeoungrang Lake,Songji Lake and HwajinpoLake.They are all originally natural and brackish waterlakes.Therefore,these lakes are not only historically veryimportant but also have high values for geological,andecological sciences.Natural lakes like lagoons are different from man-made lakes.That is,water level in natural lakes is almostconstant,cline by waterside is not much,and wet la…     

1株枯草芽胞杆菌体外拮抗6种肠道致病菌的研究

目的研究枯草杆菌BS-3株对大肠埃希菌等6种肠道致病菌的拮抗作用。方法通过在体外BS-3菌株分别与大肠埃希菌等6种致病菌混合培养后,观察不同时间内各菌的菌量变化。结果BS-3菌株与6种肠道致病菌混合培养24、48、72和96h,其菌量逐渐增加;6种致病菌的菌量随着培养时间延续逐渐减少,其中产毒性大肠埃希菌、致病性大肠埃希菌和宋内志贺菌与对照组比较差异更明显。结论BS3菌株在培养生长过程中,可抑制大肠埃希菌等6种肠道致病菌的生长。     

Time of the preovulatory LH surge in the gilt and sow relative to the onset of behavioral estrus

Two experiments were conducted to study the time of occurrence of the preovulatory LH surge in pigs. Sampling every ten minutes in six cycling gilts before and after onset of standing estrus revealed the preovulatory surge began from 8 hr before to 12 hr after the lordosis reflex was elicited. Three of six gilts initiated the preovulatory LH release coincident with the onset of estrus. Data from 28 postpartum sows, with samples drawn every six hours commencing with the onset of estrus, indicated maximum LH levels were present at the first observance of estrus. Six of the 28 sows had an LH peak 18-24 hr after the onset of estrus.     

Addendum to the Morphology of Steinernema scapterisci

This paper provides additional information on the morphology of Steinernema scapterisci. For females, the amphids are shown for the first time, and clearer scanning electron microscope (SEM) photographs of the six labial and four cephalic papillae are presented. For males, six labial and four cephalic papillae, one or two pairs of additional genital papillae, and the death shape are shown. For infective juveniles, six labial and four cephalic papillae and an elevated oral disc are shown.     

Mutagenesis of the catalytic subunit of rabbit muscle protein phosphatase-1

We have generated site-directed mutants of the catalytic subunit of rabbit muscle ppase-1. Since it is known that ppase-1 and ppase-2A are highly susceptible to inactivation by sulfhydryl reagents, we have mutagenized the six cysteine residues conserved between these two enzymes to serines. The six mutants were purified to near homogeneity by affinity chromatography on inhibitor-2-Sepharose and characterized. All six exhibited enzymatic activity. These results indicate that the catalytic mechanism of ppase-1 is different from that of the protein tyrosine phosphatases which involve a cysteinyl phosphate intermediate.     

Physiologic specialization of Colletotrichum corchori,Ikata and Yoshida,the causal organism of anthracnose of jute (Corchorus capsularis L.)

Summary Four varieties of jute,Corchorus capsularis, among fifteen screened were found to differentiate six races ofColletotrichum corchori. As regard virulence only two races, CS-1 and CS-2 among the six were of real importance.Besides recognising six races on four differential jute varieties, these were also differentiated from their cultural and morphological studies on laboratory medium as six cultural races.     

Lincomycins in the Treatment of Bacteroides Infections

Lincomycin, or the closely related derivative clindamycin, was used to treat six patients with bacteroides infection. In five of the six there was a rapid clinical response to the treatment. Lincomycin and clindamycin seem to be the antibiotics of choice for such infections.