蛋白

The immunoregulation activity of SPG-1 mainly existed in the part of sugar chain, but keeping its molecular structure intact had some effects on its immunoregulation activity; SPG sulphation had reducing effects on the immunoregulation of SPG-1, whereas, after medium and low acetification of SPG, the immunoregulation of SPG was enhanced. Conversely after high acetification, the immunoregulation of SPG-1 decreased, that is, after introducing sulfuric and acetic acid base in SPG-1, its immunoregulation changed. All of these indicated the structure of SPG-1 was closely connected with its immune activity.⑤.

甘薯糖蛋白SPG-1的免疫调节活性主要在其糖链部分，但保持分子结构的完整性对其免疫调节的活性有一定的作用；甘薯糖蛋白硫酸化对甘薯糖蛋白SPG-1的免疫调节有降低作用，而中、低度乙酰化后，可提高甘薯糖蛋白的免疫调节作用，高度乙酰化后则降低其免疫调节作用，即甘薯糖蛋白引入乙酰基和硫酸基团后，其免疫调节活性发生了改变，这些均表明了甘薯糖蛋白的结构与其免疫调节活性有密切的关系。

The main collagens in skin are type I and III. In this research, total collagens were separated from rat skin by enzymatic digest and acetum methods. The collagens were denatured at 60℃ and digested with trypsin. Characteristic peptides typical for collagen type I and III were identified with high performance liquid chromatography/mass spectrometry.

哺乳动物皮肤真皮中胶原蛋白含量约为70%，主要为是I型、III型胶原蛋白，本实验利用稀酸溶解和酶法提取了大鼠皮肤中的总胶原蛋白，将胶原蛋白粗提品在60℃变性后用胰蛋白酶进行降解，液相色谱/质谱联用法分析了两种胶原蛋白的特征多肽，利用特征多肽比较了不同生长期大鼠皮肤中I型和III型胶原蛋白相对含量。

Calretinin; fovea centralis; macula lutea; ora serrata; photoreceptors; rods; cones; optic tract; optic nerve; visual cortex; color vision; photoreception; opsin; rhodopsin; guanine nucleotide-binding protein; G protein-coupled receptors; ion channels (cyclic GMP-gated); guanylate cyclase; cyclic GMP; dark adaptation; visual pigments; polyenes; 11-cis-retinal; vitamin A; chromophores; arrestin; recoverin; phosducin; transducin; bipolar cells; retinal ganglion cells; retinal progenitor cells; amacrine cells; Mueller cells; light; retinogenesis; ommatidia; optic vesicles; retinitis pigmentosa; blindness; macular degeneration; blind spot; Mach bands; electroretinograms; binocular vision; visual acuity; vision; retina

光感受；视蛋白；视紫质；鸟苷酸-结合蛋白；G蛋白-电偶受体；离子通道；鸟苷酸环化酶；环鸟苷酸；暗适应；视色素；多烯；11 - cis -视网膜的；抗干眼醇；发色团；抑制蛋白；恢复蛋白；phosducin；转导蛋白；双极细胞；视网膜神经节细胞；视网膜祖细胞；无长突细胞；米勒细胞；光；retinogenesis；小眼；视泡；色素性视网膜炎；盲的；黄斑变性；盲点；马赫带；视网膜电流图；双目视觉；视敏度；视觉；视网膜

The family of glycophosphoproteins comprising osteopontin, bone sialoprotein, dentin matrix protein 1, dentin sialophosphoprotein and matrix extracellular phosphoglycoprotein — small integrin-binding ligand N-linked glycoproteins — are emerging as important players in many stages of cancer progression.

糖磷酸化蛋白包含骨桥蛋白，骨唾液蛋白，牙质基质蛋白及细胞外基质糖蛋白――小的整联蛋白结合配体 N 联结糖蛋白――作为癌症演进早期的一个重要物质。

It was found that La〓 does not affect the binding affinity between calmodulin and Polistes Mastoparan, a known calmodulin binding peptide, neither the conformation of the ternary complex. Excessive amount of La〓 result in the decrease of the binding constant and the disrupted conformation. The binding affinity of La〓 to calmodulin increased in the ternary complex (La〓-CaM-Mas/Mas X), which suggested the coordination between the two global domains. The binding priority between the two global domains is also changed: La〓 more is likely to bind to the C-terminal of calmodulin than to N-terminal, thus facilitates the binding of Mas/Mas X to the C-terminal of calmodulin as the first step of the Mas/Mas X binding.

在以钙调蛋白结合肽—Polistes Mastoparan和Mastoparan X为对象的研究中，我们发现，除非过量，否则La〓的存在不影响钙调蛋白与Mas间的结合常数以及构象；过量的La〓（La〓/CaM摩尔比＞4）将引起钙调蛋白结合功能的下降，同时明显改变金属-钙调蛋白-Mas三元复合物的构象；La〓对Ca〓CaM-Mas的N末端表现高度选择性，显示了在三元复合物中两种离子间的协同效应；在三元体系中，La〓与钙调蛋白的亲和力明显上升，而且亲和力上升的程度因钙调蛋白结合肽的不同而有明显差异，显示了作用的选择性；Mas和MasX的存在改变了La〓在钙调蛋白上的结合顺序，La〓很可能优先与C末端结合，从而使Mas/Mas X首先与C末端结合，该顺序与钙离子相同；La〓的参与使三元复合物在动力学上更加稳定。

Results:(1) The conchosporangia of brown yellow mutant was smaller and mature time was 15 days later than the parents. Thallus grew slowly in early stage, but average daily growth rate reached (7.50±1.18) cm after blade length was over 60 cm.(2)The emerald conchocelis were easy to mature and had special developmental mode that the spherulocytes can directly develop into sporangial branchlets and conchosporangia without thick conchocelis stage. Emerald blades were low in RPE with just (6.4710±0.0184) mg/g dry mass. The thallus grew quickly, average daily growth rate reached (11.95±2.33) cm after blade length was over 60 cm.(3) The conchocelis filaments of breen mutant were thin and short. Thallus grew slowly and had low contents of three phycobiliprotenin and chlorophyl.

实验进行50d。结果：(1)褐黄色突变体藻蓝蛋白和别藻蓝蛋白含量低；孢子囊枝的细胞较小，且大量形成时间比亲本晚15d；幼苗培养初期日平均生长量仅为(1.22±0.28)cm，当叶片长到60cm左右时生长优势逐步凸显，日平均生长量可达(7.50±1.18)cm；(2)翠绿色丝状体容易成熟，发育方式特殊，营养藻丝不经过藻丝加粗阶段，直接由球形细胞发育成孢子囊枝和壳孢子囊；翠绿色叶状体藻红蛋白含量低，仅有(5.5130±1.0496)mg/g；叶状体生长快速，60cm长的藻体日平均生长量高达(11.95±2.33)cm；(3)褐绿色突变体藻蓝蛋白、别藻蓝蛋白和藻红蛋白这3种色素蛋白和叶绿素的含量均较低；藻丝细胞短且细，叶状体生长速度较慢。

By use of site mutation strategy and PCR technology, we obtained the gene P12X3C that includes full length P1, 2A, 3C and a part of 2B and 3B and the gene P12X3C3D that includes full length P1, 2A, 3C, 3D and a part of 2B and 3B. After being digested by restriction enzyme respectively, the gene P12X3C and the gene P12X3C3D were cloned into the pcDNA3. 1 and pTARGET expression vector that were digested by the same enzyme. Recombinant plasmids were checked by restriction enzyme analysis and nucleic acid sequencing. Further more, recombinant plasmids were transfected into BHK-21 cells by using lipoid. The proteins of foot-and-mouth disease virus , which were expressed in BHK-21 cells, were confirmed by sandwich-ELISA and fluoroscopy, and the capsid of FMDV was tested by electron microscope. In order to evaluate enhanced immune response of guinea pigs against FMDV, DNA vaccines which were designed to produce viral capsids lacking infectious viral nucleic acid and contained the gene P12X3C and the gene P12X3C3D were injected respectively with FMDV 3D protein which was expressed in Pichia Pastoris Secreted expression System and purified or with pcDNA3. 1/IFN which includes the gene IFN-α of cattle. Subsequently, Recombinant plasmids were injected to cattles with or without pcDNA3. 1/IFN. Anti-FMDV antibodies were detected by ELISA, and the T lymphocyte proliferation response was tested by MTT assay, neutralization antibodies titers were analyzed by micro-neutralization assay.

为研制带有O型口蹄疫病毒(Foot-and-Mouth Disease Virus，FMDV)China99株结构蛋白基因及多个非结构蛋白基因的DNA疫苗，本研究通过定点突变方法和PCR扩增方法，获得包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C以及部分2B、3B编码基因的片段P12X3C和包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C、3D以及部分2B、3B编码基因的片段P12X3C3D，将获得的基因片段直接/酶切后与同样处理的真核表达质粒连接，分别得到重组质粒pcDNA3.1/P12X3C和pcDNA3.1/P12X3C3D、pTARGET/P12X3C3D；对重组质粒进行序列测定、分析，并将重组质粒分别转染BHK-21细胞，通过双抗体夹心ELISA方法和间接免疫荧光标记方法检测细胞中FMDV抗原的表达，用电子显微镜观察病毒空衣壳的组装；为评价重组质粒作为DNA疫苗对实验动物及本动物的免疫效果，将重组质粒经肌肉注射方法接种豚鼠，并与酵母表达的纯化FMDV China99株3D蛋白及带有牛α干扰素的真核表达质粒pcDNA3.1/IFN分别/同时免疫，第二次免疫后第三周豚鼠攻以1OOID〓或1000ID〓的O型FMDV China99株；随后将质粒pcDNA3.1/P12X3C、pcDNA3.1/P12X3C3D与带有牛α干扰素的真核表达质粒pcDNA3.1/IFN同时免疫牛，三周后经牛舌皮攻以10〓ID〓的O型FMDV China99株。

By use of site mutation strategy and PCR technology, we obtained the gene P12X3C that includes full length PI, 2A, 3C and a part of 2B and 3B and the gene P12X3C3D that includes full length PI, 2A, 3C, 3D and a part of 2B and 3B. After being digested by restriction enzyme respectively, the gene P12X3C and the gene P12X3C3D were cloned into the pcDNA3.1 and pTARGET expression vector that were digested by the same enzyme. Recombinant plasmids were checked by restriction enzyme analysis and nucleic acid sequencing. Further more, recombinant plasmids were transfected into BHK-21 cells by using lipoid. The proteins of foot-and-mouth disease virus, which were expressed in BHK-21 cells, were confirmed by sandwich-ELlSA and fluoroscopy, and the capsid of FMDV was tested by electron microscope. In order to evaluate enhanced immune response of guinea pigs against FMDV, DNA vaccines which were designed to produce viral capsids lacking infectious viral nucleic acid and contained the gene P12X3C and the gene P 12X3C3D were injected respectively with FMDV 3D protein which was expressed in Pichia Pastoris Secreted expression System and purified or with pcDNA3.1/lFN which includes the gene IFN-a of cattle. Subsequently, Recombinant plasmids were injected to catties with or without pcDNA3.1/IFN. Anti-FMDV antibodies were detected by ELISA, and the T lymphocyte proliferation response was tested by MTT assay, neutralization antibodies liters were analyzed by micro-neutralization assay.

为研制带有O型口蹄疫病毒(Foot-and-Mouth Disease Virus，FMDV)China99株结构蛋白基因及多个非结构蛋白基因的DNA疫曲，本研究通过定点突变方法和PCR扩增方法，获得包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C以及部分2B、3B编码基因的片段P12X3C和包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C、3D以及部分2B、3B编码基因的片段P12X3C3D，将获得的基因片段直接／酶切后与同样处理的真核表达质粒连接，分别得到重组质粒pcDNA3.1／P12X3C和pcDNA3.1／P12X3C3D、pTARGET／P12X3C3D；对重组质粒进行序列测定、分析，并将重组质粒分别转染BHK-21细胞，通过双抗体夹心ELISA方法和间接免疫荧光标记方法检测细胞中FMDV抗原的表达，用电子显微镜观察病毒空衣壳的组装；为评价重组质粒作为DNA疫苗对实验动物及本动物的免疫效果，将重组质粒经肌肉注射方法接种豚鼠，并与酵母表达的纯化FMDV China99株3D蛋白及带有牛α干扰素的真核表达质粒pcDNA3.1／IFN分别／同时免疫，第二次免疫后第三周豚鼠攻以100ID_(50)或1000ID_(50)的O型FMDV China99株：随后将质粒pcDNA3.1／P12X3C、pcDNA3.1／P12X3C3D与带有牛α干扰素的真核表达质粒pcDNA3.1／IFN同时免疫牛，三周后经牛舌皮攻以10~4ID_(50)的O型FMDV China99株。

After amplying a 2.2kb fragment form the PPV-SC1 RF-DNA,we clone the fragment into pMD 18-T,named pTNSl.The whole sequence which is 1989 bp long was determined by sequencing, including the complete ORF of PPV-SC1 NS1 which encoding 662 amino acids.Alignment of pairs of sequence indicates that there are 98% and 99% similary with other porcine parvovirus strains Kresse and NADL-2, respectively. Multiple sequence alignment discloses that there are a few difference between ppv-scl nsl gene and other ppv nsl gene: A-G at 39nt,T-C at 153nt,A-G at 175nt, A-C at 1117nt, A-C at 1535nt .Alternative codon in ppv-scl nsl have distinctly different frequentfy by codonbias analysis at EMBOSS(http://genopole.toulouse.inra.fr/bioinfo/emboss). Thereis not distinct hydrophobicity and transmenbrane helices in ppv-scl nsl protein. Struction domain anslysis of PPV-SC1 NS1 protein indicate that there are a ATP/GTP-binding site motif A at 398-405,16 Protein kinase C phosphorylation site,21 Casein kinase II phosphorylation site,and 3 cAMP/cGMP-dependent protein kinase phosphorylation site.At the same time ,there is a same motif between ppv-scl nsl and Poxvirus D5 protein-like which may share in the same fuction which is necessary during virion duplication.

将PPV-SC1 NS1序列与其他PPV NS1基因进行多序列比对，结果显示，PPV-SC1 NS1与其他的PPV NS1的同源性较高，仅存在个别的差异，分别是第39位A→G，第153位T→C，第175位A→G，第1117位A→C，第1535位A→C；同源搜索比较表明，PPV-SC1与PPV NS1同源性可达98％、99％，与其他的细小病毒NS1基因也存在很大的保守性；密码子偏向性分析结果表明PPV-SC1 NS1基因在同一氨基酸的不同密码子的选择上存在一定的偏向性；PPV-SC1 NS1蛋白总体上说具有亲水性不存在明显的疏水性区段，用swiss TMPRED软件预测PPV-SC1 NS1的跨膜区，返回的结果并没有得到有显著意义的跨膜区的存在；根据基于motif数据库的结构域预测，PPV-SC1 NS1的第393-415位氨基酸残基存在潜在的ATP／GTP结合位点，该蛋白还存在16个蛋白激酶C磷酸化位点，21个酪蛋白激酶2磷酸化位点，3个cAMP-／cGMP依赖蛋白激酶磷酸化位点，PPV-SC1 NS1蛋白与POX_D5(痘病毒D5蛋白)具有一致的保守结构域，推测NS1可能与POX_D5有类似的功能。

The results indicated that the well-resolved and reproducible 2-DE pattern of Golgi apparatus isolated from gastric cancer cells was obtained , and twelve proteins were identified, including proteins related to protein synthesis and folding, membrane fusion proteins, regulatory proteins, transportation proteins, proteins related to apoptosis, proteins related to cell proliferation.

结果显示分离出的纯度较高的高尔基体建立了分辨率和重复性均较好的双向电泳图谱，运用质谱技术鉴定出12个蛋白质，包括蛋白合成相关蛋白，膜融合蛋白，调节蛋白，凋亡相关蛋白，运输蛋白，细胞增殖分化相关蛋白。

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