酶蛋白

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。

The esterase isozyme in leaf at acicula forming stage and the peroxidase isozyme in peanut leaf at pod producing stage was related to the formation of coarse protein and coarse fattiness of seed.

下针期花生叶片的酯酶同工酶与结荚期叶片过氧化物酶同工酶与其果仁粗脂肪、粗蛋白的形成有关；饱果期花生叶片和果仁酯酶同工酶与花生的单株果数和果重相关，并且对花生果重、仁重有一定的影响。

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

Result s:10 μmol/L retinoids could inhibit the Bel-7402 proliferation and decrease the colony forming of liver cancer cells on soft agar significantly.So some biochemical markers indicating the proliferation status of liver cancer cells changed significant ly, including the significant activity increase of ornithine carbamyl transferase, tyrosine-α-ketoglutaric transaminase and alkaline phosphatase, the significant decrease α-fetoprotein secretion,γ-glutamyltranspetidase and aldolase activity.

结果：10μmol/L维甲酸显著抑制肝癌细胞增殖，并使肝癌细胞软琼脂集落形成率明显减少，使代表肝细胞分化的酶鸟氨酸氨基甲酰转移酶、酪氨酸-α-酮戊二酸转氨酶和碱性磷酸酶比活力明显升高，甲胎蛋白分泌量、γ-谷氨酰转肽酶和醛缩酶比活力明显下降。

PSE7 and SOJB had the similar binding proteins in tobacco, suggesting that they may share some similar biological functions. The ubiquitin-conjugating protein and peptidase involved in protein degradation were found binding with SOJB but not PSE7. Both SOJB and PSE7 could bind with transmembrane transport protein, SOJB bound with copper transporter protein, and PSE7 bound with water channel protein. The results suggested that the target of SOJB and PSE7 may be located in cytoplast.

PSE7和SOJB在烟草中存在几个相似的结合蛋白，表明二者具有共享的生物学功能；SOJB能与肽酶和泛素结合蛋白（与蛋白质降解有密切关系）互作，而PSE7没有筛选到类似蛋白；SOJB和PSE7均能与跨膜运输蛋白结合，PSE7与水通道蛋白结合，SOJB与铜离子转运蛋白结合，表明二者在胞质内均存在作用靶标。

The types of histone methyltransferases, the relationship between methylation of Lysine 9 of H3 and the formation of heterochromatin, gene regulation in euchromatin, and that with DNA methylation, were mainly introduced.

主要阐述了组蛋白甲基转移酶的类型，组蛋白H3中第9位赖氨酸甲基化与异染色质的形成、常染色体中基因表达的调控，以及与DNA甲基化之间的关系，说明了组蛋白甲基化与组蛋白乙酰化、磷酸化的相互关系，指出组蛋白甲基化对维持细胞各种状态的平衡起到极其重要的作用。

This article focus on enzyme gene clone and heterogenesis expression,site directed modify and orthogenesis,fu-sion protein and fusion enzyme,enzyme mimics(abzyme and molecular imprinting)and telomere .

本文着重介绍了酶基因克隆与异源表达、酶分子的定向改造和进化、融合蛋白与融合酶、酶的人工模拟和端粒酶，综述了分子酶工程的研究进展、趋势及其应用。

Comparing with chicken egg-white lysozyme which is widely used in clinic, hLYZ has many advantages: it is a natural protein in human body, so it is safe and harmless when used as a drug; its bioactivity to kill bacterium is three times higher than chicken egg-white lysozyme, and its thermal stability is much higher, too; it also has many other specific important functions which are not related with its catalysis, such as anti-virus and anti-tumor and improving immunization.

人溶菌酶与目前临床上应用最多的鸡蛋清溶菌酶相比具有以下优势：人溶菌酶是人体内的蛋白，具有天然的相容性，没有毒性和副作用；人溶菌酶比鸡蛋清溶菌酶的杀菌活性高3倍，而且热稳定性也要远高于鸡蛋清溶菌酶；人溶菌酶还有多种特有的与其自身的催化作用无关的理化作用，如抗病毒和抗肿瘤。

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。

This one mode pays close attention to network credence foundation of the businessman very much.

这一模式非常关注商人的网络信用基础。

Cell morphology of bacterial ghost of Pasteurella multocida was observed by scanning electron microscopy and inactivation ratio was estimated by CFU analysi.

扫描电镜观察多杀性巴氏杆菌细菌幽灵和菌落形成单位评价遗传灭活率。