表达

The highest expression level of AtACS5::GUS fusion was detected in the 3-d-old etiolated seedlings, and the developmental expression profile of AtACS5 was not affected by the etr1-1 mutation.

在转基因拟南芥的整个生活周期中，AtACS5：：GUS和AtACS7：：GUS融合基因都具有相似的表达谱；待外源乙烯处理后，光照生长下的2周幼苗中AtACS7：：GUS的表达显著增加，而AtACS5：：GUS的表达不受乙烯处理的影响。3天苗龄的转基因黄化苗中，AtACS5：：GUS具有很高的表达水平，并且ETR1-1基因突变对AtACS5：：GUS在生长发育过程中的表达模式没有明显的影响。

Methods:Twenty-one gastric carcinoma tissues and correponding non-cancer mucosae were examined by in situ hybridization with digoxigenin-labeled nm23-H1cDNA probes.

此后，在多种动物模型或人类某些实体癌中发现nm23-H1基因表达水平与肿瘤转移呈负相关［2～5］，但少数作者的实验结果表明，nm23-H1基因与转移无关［6，7］。nm23-H1基因在胃癌中表达仅见少数报道［7，8］，结果不一，本组实验应用地高辛标记cDNA探针和免疫组化方法检测石蜡包埋组织中nm23-H1基因及其表达产物nm23/NDPK在胃癌中的表达，探讨nm23-H1基因表达与胃癌的临床病理关系。

The results of in situ hybridization showed that the gene of Mdde was expressed mainly in epidermis of the body wall and fat bodies, and no expression signal was detected in tracheae, intestine and musculature. The gene of Mdde in epidermis expressed both in challenged larva and no-challenged larva, whereas the gene of mdde in fat body expressed only in challenged larva.

原位杂交的结果表明：Mdde基因在家蝇幼虫体内主要表达于表皮和脂肪体，在气管、肠和肌肉组织中没有检测到Mdde基因的表达；表皮中Mdde基因在微生物刺激前后均有表达，而脂肪体中Mdde基因经微生物刺激后才表达，刺激前不表达。

Results VEGF-C and VEGFR-3 are all expressed on both mesenchymal cells and vascular endothelium of the embryos during early period. In middle and latter period, VEGFR-3mRNA is mainly expressed on cell plasm of alimentary canal, but it has no expression on vascular and lymphatic endothelium; The expression of VEGFR-3 is extensive and positive expression is seen on the vascular and lymphatic endothelium.

在胚胎中、后期消化道组织中VEGF-C主要表达于消化管壁组织细胞以及肠系膜内，在血管和淋巴管内皮细胞则无表达；VEGFR-3亦表现为广泛的表达，于血管和淋巴管内皮细胞可见阳性表达，并随着胚胎的发育，其在淋巴管内皮细胞的阳性表达率明显增加。

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。

In the aspect of research and development of HAS, the United States, Japan, united Kingdom, France and Italy recently tried tk use lower eucaryote, espesially yeast expression system to express HAS, obtained outstanding progress.

HSA的表达及工程化：将含HSA基因的酵母转化子单个菌落分别接种，并且分别进行甲醇诱导表达；将酵母表达的HSA蛋白质经SDS-PAGE电泳鉴定，筛选出多拷贝整合和高水平表达HSA菌株；将高表达HSA工程菌加保护剂后分别在液氮和－70C°低温冰箱内保存。

The correlation between Survivin and p53 expression and histological tyes and clinical stages in ovarian carcinoma were analysed.

结果： 1。卵巢癌p53和Survivin蛋白阳性表达率在卵巢癌组织中P53蛋白阳性表达率为54.84%(34/62)与正常对照组比较有显著差异(p＜0.01)；在浆液性囊腺癌中P53蛋白表达阳性表达率为56.52%(26/46)在黏液性囊腺癌中P53蛋白阳性表达率为57.14%(8/14)两组间无明显差异(p＞0.05)。

When aborted rats were injected with 10 IU/g IFN-γin group D, compared with group A, expression of TNF-αinmmuopositive substance in the nucleus pre-opticus medialis, nucleus supra-opticus, nucleus pre-opticus mango celluaris, nucleus periventricularis hypothalami, nucleus paraventricularis hypothalami, nucleus arcuatus hypothalami were highly increased, conversely, the ones in the nucleus pre-opticus suprachiasmaticus, neurohypophsis, adenohypophysis, ovary were obviously decreased, the change in the uterus was not remarkable; Compared with group B, expression of TNF-αin the arcuatus hypothalami was obviously enhanced, while in the nucleus pre-opticus suprachiasmaticus, nucleus pre-opticus mango celluaris, nucleus periventricularis hypothalami, nucleus paraventricularis hypothalami, nucleus ventromedialis hypothalami, nucleus dorsomedialis hypothalami, neurohypophsis, pars intermedia, ovary, the results were on the contrary, the uterus had no significantly variation; Compared with group C, expression of TNF-αin the nucleus pre-opticus medialis, nucleus supra-opticus, nucleus paraventricularis hypothalami, nucleus arcuatus hypothalami, adenohypophysis and pars intermedia had an obviously risen trend, however, the variation in the nucleus pre-opticus suprachiasmaticus, nucleus periventricularis hypothalami, neurohypophsis, pars intermedia, follicle and corpus luteum was completely opposite, uterus had no obviously change.

外源腹腔注射10 IU/g IFN-γ后，同A组相比：D组TNF-α免疫阳性产物在下丘脑视前内侧核、视上核、视前大细胞核、室周核、室旁核、弓状核显著增高，在视交叉上核显著降低，在其它核团较A组无显著差异，在神经垂体、垂体前叶、卵巢中阳性产物表达均较A组显著降低，在子宫中的表达较A组变化不明显；同B组相比：D组TNF-α免疫阳性产物在弓状核显著增高，在视交叉上核、视前大细胞核、室周核、室旁核、腹内侧核、背内侧核、神经垂体、垂体中间部、卵巢各部阳性物质表达均显著降低，在子宫表达变化不显著；同C组相比：D组下丘脑视前内侧核、视上核、室旁核、弓状核、垂体前叶与中间部均显著增高，在视交叉上核、室周核、神经垂体、卵泡、黄体中均显著降低，子宫中表达变化不明显。

Results: In cellular soluble protein electrophoresis,specific band about 75 KDa was observed in 593 strain.The differences of protein expression at 85,60,50,40KDa between 593 strain and 18 strain were observed.In extracellular soluble protein electrophoresis,specific bands at 75 KDa and 40 KDa were observed in 593 strain;specific band at 50 KDa was observed in 18 strain.The difference of protein expression amount in about 60KDa between 593 strain and 18 strain was observed.

结果：菌体可溶性蛋白电泳图中见593号菌株在75 KDa左右存在特异表达的蛋白条带，且二者在85、60、50、40 KDa左右蛋白表达量上存在差异；菌外可溶性蛋白电泳图中见593号菌株在75、40 KDa左右有特异表达的蛋白条带，而18号菌株在50 KDa左右有特异表达的蛋白条带，二者在60 KDa左右蛋白表达量上存在差异。

What are your goals and strategies for growth?

你的成长目标和策略是什么？

And unto the angel of the church in Sardis write; These things saith he that hath the seven irits of God, and the seven star I know thy works, that thou hast a name that thou livest, and art dead.

3：1 你要写信给撒狄教会的使者，说，那有神的七灵和七星的，说，我知道你的行为，按名你是活的，其实是死的。