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Ultrastructural observations on human spermatozoa preserved by 3% glutaraldehyde and prepared by freeze-etch replica techniques were carried out.The results were as follows: 1.According to the structural features,the head of human sperm may be divided into three distinct different regions,namely,fore-head,mid-head and hind-head.This regional distinction may represent the functional differences. 2.Regional difference of the intramembranous particles of the inner-and outer- leaflets of the plasma membrane was...

Ultrastructural observations on human spermatozoa preserved by 3% glutaraldehyde and prepared by freeze-etch replica techniques were carried out.The results were as follows: 1.According to the structural features,the head of human sperm may be divided into three distinct different regions,namely,fore-head,mid-head and hind-head.This regional distinction may represent the functional differences. 2.Regional difference of the intramembranous particles of the inner-and outer- leaflets of the plasma membrane was also revealed.The intramembranous particle is very rich in the fore-head region of inner-leaflet.However,the mid-bead region is very smooth and there are only a few particles attached on its fractured face,but groups of extrinsic membrane particles can be seen on its inner plasma-face.As for the hind-head region the inner-and outer-leaflets of the unit membrane contact closely with each other and attach tightly to the nuclear membrane.It is suggested that this structure is prerequisite for attaching the tail to the head firmly.The PF-face of sperm tail pla- sma membrane is rich in particles but without special pattern in arrangement. 3.The acrosome is extremely large,almost 8/10~9/10 of the nuclear surface is covered by it,its posterior margin sometimes is described as “posterior ring”.Two distinc- tive parts may be seen on the acrosome:The fore-acrosome,which corresponds to the fore-head region,is rich in membrane particles.It looks plump in appearance and is filled up with content.There is also pore-like structure at the internal membrane of the acrosomal cap.The hind-acrosome part,corresponds to the mid-head region.There are less particles than the fore-acrosome part.Its external and internal membranes contact with each other and it looks lean in features. 4.The polarity of the nucleus is very obvious.On the anterior part of the nucleus the nuclear pores are distributed irreglarly,and in the middle part there is no pore, but in the posterior part the nuclear pores are large and arranged closely as regular hexagonal arrays.A hypothesis is proposed that the anterior part of the nucleus may regulate the activity of acrosomal hydrolase by chemical message which is presumably produced by the chromosome and transfered from the nuclear pore through the pore of the acrosomal membrane into the cavity of the acrosome.The posterior part of the nucleus is thought to be the place where the chemical message,which may be the pro- duct of a particular gene,is transfered through the nuclear pores to the tail,and it can adiust the absolutely synchronous movement of the axial microfilarnents and modulate the oxidation phosphorylation and energy-release of the mitochondria.

应用冷冻复型电子显微镜技术,研究了正常人类精子的超微结构,通过观察分析,作者发现如下现象并提出了相应的推论。一、按照结构特点,精子头可分为前、中、后三头区,形态结构的这种区域性,反映了生理功能的区域性。二、质膜内外片的膜内颗粒,具有不同的区域性;在前头区内片膜内颗粒丰富;中头区内片从断裂面看膜面平滑、颗粒少,从胞质面看有成群散在的表在性膜蛋白粒;后头区质膜内外片贴附紧密,并与核膜相贴,结构坚韧,可提供精子尾保持牢固附着的部位;精子尾质膜 PF 面颗粒丰富,但未见任何特异的排列方式。三、顶体极大,分前后两部,遮盖核表面十分之八九;前顶体位于前头区范围,膜内颗粒丰富,外形肥厚,充满内容物,内膜层有孔状结构;后顶体则位于中头区范围,膜内颗粒少,内外膜相贴,外形干瘪。四、细胞核极性明显,在前头区核孔无规律,中头区核孔不明显,后头区核孔大而密,排列成规律的六角式;作者推论:核的前头区可能发出控制顶体囊蛋白水解酶活性及细胞识别的化学信息,经核孔传出,后头区大概也发出化学信息,经核孔输向尾侧,以协调统筹轴丝微丝运动的绝对同步及线粒体的能量释放。

BALB/c mice were repeatedly inoculated subcutaneously with purified rat source Pneumocystis carinii (P.c.) cysts.Six weeks later,the spleen cells were fused with NSI myeloma cells,and a mouse hybridoma producing monoclonal antibody against P.c. was establishted and cloned,which was named as 4D7 McAb.Its mean of chromosome was 82.The 4D7 McAb was shown to be IgG1 subclass.It reacted mainly with 54 kDa polypeptide of P.c. in EITB.4D7 McAb could ...

BALB/c mice were repeatedly inoculated subcutaneously with purified rat source Pneumocystis carinii (P.c.) cysts.Six weeks later,the spleen cells were fused with NSI myeloma cells,and a mouse hybridoma producing monoclonal antibody against P.c. was establishted and cloned,which was named as 4D7 McAb.Its mean of chromosome was 82.The 4D7 McAb was shown to be IgG1 subclass.It reacted mainly with 54 kDa polypeptide of P.c. in EITB.4D7 McAb could clearly recognize P.c.by immunohistochemical staining in the specimens from confirmed Pneumocystis carinii pneumonia(PCP)patients,and the sensitivity and specificity were both 100% for diagnosis of PCP(11/11).No cross reaction of 4D7 McAb was found with Pneumococcus,Candida albicans and Cryptococcus neoformans,as well as all examined normal human tissues.

从感染肺孢子虫肺炎大鼠模型中分离纯化肺孢子虫包囊并免疫BALB/c小鼠,采用杂交瘤技术,获得1株能长期稳定分泌抗人源性肺孢子虫单克隆抗体杂交瘤细胞株4D7。其染色体众数为82,分泌IgG1亚型抗体。该单抗可识别54kDa肺孢子虫包囊蛋白区带,并能准确识别肺孢子虫包囊,其灵敏性与特异性可达100%(11/11);并与正常人体组织、肺炎双球菌、白色念珠菌和新型隐球菌无交叉反应。

Soluble antigens extracted from cyst wall(CW),cysttozoites (CT) and cyst fluid(CF) of Sarcocystis fusiformis and S.gigantea from naturally infected animals were analysed for the molecular weights and immunological characteristics by SDS—PAGE.Western blot and ELISA.Bands from 10 to 26 with Mwranging from approximately 9 to 100kDa were separated from various antigenic extracts.Antigens from CW and CT are mainly larger molecular proteins,while the antigens from CF mainly consist of smaller molecular proteins...

Soluble antigens extracted from cyst wall(CW),cysttozoites (CT) and cyst fluid(CF) of Sarcocystis fusiformis and S.gigantea from naturally infected animals were analysed for the molecular weights and immunological characteristics by SDS—PAGE.Western blot and ELISA.Bands from 10 to 26 with Mwranging from approximately 9 to 100kDa were separated from various antigenic extracts.Antigens from CW and CT are mainly larger molecular proteins,while the antigens from CF mainly consist of smaller molecular proteins under 50kDa.ACA54 ultragel chromatography was adopted to isolate and purify specific antigens from crude antigen preparation of S.fusiformis cysts.72kDa protein from purified antigens and 175,53,72,80,90,and 100kDa proteins from whole cyst extracts were identified by SDS—PAGE and Western blot.It has been demonstrated by ELISA that the purified antigen had lower sensitivity and higher specificity in comparison with the crude antigens.

本文对采用十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS—PAGE)对梭形住肉孢子虫和巨型住肉孢子虫的包囊壁(CW)、包囊液(CF)与缓殖子(CT)三部分的可溶性蛋白进行了分析,从各组份区分出10—26条蛋白带,分子量9—100kDa,CW以高分子量蛋白为主,CF则以50kDa以下低分子量蛋白带居多,CT各种蛋白带分布较均匀。应用超凝胶ACA54柱层析和SDS—PAGE对梭形住肉孢子虫包囊蛋白进行了分离纯化,分离出72kDa蛋白,经免疫印渍术鉴定为单特异性抗原、经对86头水牛的ELISA检疫分析,其纯化抗原较未纯化的粗抗原,特异性增强,但敏感性降低。

 
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