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31.
猪带绦虫六钩蚴cDNA文库的构建与筛选 总被引:1,自引:0,他引:1
在体外将猪带绦虫虫卵孵化为有活力的六钩蚴。采用商品化试剂盒抽提六钩蚴总 RNA、m RNA,反转录为双链c DNA,再加 Eco R Adapter。在去除小片段后 ,dsc DNA与 λgt11噬菌体 DNA连接 ,经蛋白包装 ,构建了猪带绦虫六钩蚴λgt11c DNA文库。该文库效价为 3.5× 10 9pfu/ m L ,重组 DNA片段大小为 0 .5~ 3.2 kb,平均为 1.6 kb,蓝白斑比 1∶ 9。用抗体探针对文库进行免疫学筛选 ,在消除非特异性反应的基础上筛选约 10 6 重组子 ,共得到 118株强阳性克隆 ;应用 PCR鉴定上述部分阳性克隆 ,均扩增到 0 .5 kb以上的片段。结果显示 ,构建的文库合格 ,含六钩蚴所有抗原基因 ,可用于六钩蚴 c DNA克隆的筛选 ;用免疫学与 PCR联合筛选 c DNA文库 ,可消除假阳性 相似文献
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Mouse pronuclear stage embryos with intact slit zona pellucida (manipulated) were cultured in vitro until the hatched blastocyst stage in simplex optimized medium with higher K+ concentration (KSOM) prepared with three different water types: tap, deionized reverse osmosis (D‐O) water and Milli‐Q system (M‐Q) water. The culture media were supplemented with or without protein and ethylenediaminetetraacetic acid (EDTA, disodium salt). The rates of hatched blastocysts were significantly affected (p < 0.01) by micromanipulation, protein supplement and water source. The water source has no influence (p > 0.05) on development in EDTA‐supplemented protein‐free culture media, whereas in EDTA‐ and protein‐free culture media, the water quality significantly (p < 0.001) affected the rates of development, with higher rates in media prepared with M‐Q water. The micromanipulated embryos showed higher sensitivity to the water quality (p < 0.01). It worth mentioning that the rates of hatched blastocysts in protein‐free culture media were very low (0–7.5%). Furthermore, the three different water types were analysed by measuring the electrical conductivity, inorganic ions, total organic carbon and endotoxins to evaluate the purity. M‐Q water showed the lowest levels of inorganic ion, total organic carbon and endotoxin concentrations. We concluded that manipulated mouse embryos are good system to evaluate the quality of water used in biological system. 相似文献
34.
分析了核对、打孔、光照透明、光电机器阅读和扫描等几种快速手工和自动阅卷评分方法。采用我校目前广泛使用的“成绩记载”中的浮动“阅卷面板”鼠标点击,以及自动阅读机与“成绩记载”间的“数据转换”程序,可大大提高成绩录入、成绩分析和试题分析速度。 相似文献
35.
3种农药的微核效应研究 总被引:6,自引:0,他引:6
运用蚕豆根尖微核技术研究3种农药(甲胺磷、甲基硫菌灵和盐酸吗啉胍)在不同浓度下的诱变效应。结果表明,3种农药诱发蚕豆根尖细胞微核率与对照组相比均有不同程度的上升(P<0.05或P<0.01),农药浓度与微核率呈剂量-效应关系,盐酸吗啉胍的诱变效应大于其他2种农药。 相似文献
36.
37.
Y Maruyama Y Suzuki A Kazusaka S Fujita 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2001,63(5):493-497
The intracellular dynamics of norsalsolinol, a neurotoxin candidate causing parkinsonism-like symptoms, in PC12 cells was studied. We found that dopamine and norsalsolinol are co-localized to secretory granule layer by sucrose density gradient in norsalsolinol-treated PC12 cells. The norsalsolinol was actively taken up into isolated secretory vesicle fraction from PC12 cells with a Km value of 41.5+/-6.8 microM. The uptake of 10 microM of norsalsolinol was sensitive to reserpine (1 microM), an inhibitor of vesicular dopamine transporter, and dopamine, an endogenous substrate, but insensitive to GBR-12909, an inhibitor of dopamine transporter on plasma membrane. In norsalsolinol-treated PC12 cells, exposure to high K+ or ATP resulted in simultaneous release of norsalsolinol and dopamine. Time course of a release of dopamine and that of norsalsolinol evoked by 50 mM KCl or 100 microM ATP corresponded to each other. These releases were dependent on the concentrations of secretagogues. These data suggest that norsalsolinol is taken up with dopamine into secretory vesicle via vesicular catecholamine transporter. 相似文献
38.
I.-S. Park G. C. Choi D. S. Kim Y. K. Nam 《Journal of the World Aquaculture Society》2002,33(4):494-500
For the evaluation of hormonal control of spermiation in fish, a method to quanify the spermiation response of mature Rhynchocypris oxycephalus (Sauvage and Dabry) to hormonal therapy is described. Spermatocrit was determined after 7 min centrifugation at 18,000 ± g and sperm density was estimated by a standard hemocytomer method. Sperm density can be predicted from spermatocrit since their relationship is linear as described by the regression equation, Y = 3.68X - 27.18 ( R 2 = 0.82, N = 50). where Y is spermatocrit and × is sperm density. Milt production by mature R. oxycephalus was highest at 24 h after injection of 1,000 IU human chorionic gonadotropin (HCG) and 50 μg luteinizing hormone-releasing hormone analogue (LHRHa) per kg body weight. Increased milt production coincided with low spermatocrit and sperm density levels. These results demonstrate that spermiation in mature R. oxycephalus can be reliably evaluated by a spermatocrit method and that HCG and LHRHa are effective in stimulating of spermiation in this species. 相似文献
39.
Huang Yahong FANG Xian-chu LIANG Wanqi FANG Pingyi TANG Li CHEN Jian-xiu ZHANG DaBing 《水稻科学》2002,10(2)
Genetic variation, including agronomic trait variation, often occurs in somatic cell culturing. In this study, we compared the main agronomic traits of two rice mutants, M3 and M14, which were derived from Shenxiangjing 5 somatic cell culturing. Significant differences were found between the two mutants and the wild rice Shenxiangjing 5 (Table 1). Results were as follows: 相似文献
40.
Disappearance of acetic acid from the bovine reticulorumen at basal and elevated concentrations of acetic acid. 总被引:2,自引:0,他引:2
Disappearance of acetic acid was quantified to determine whether removal of this acid from the reticulorumen is altered when ruminal acetic acid concentrations are elevated. Ruminally fistulated beef steers (n = 3 per experiment; BW = 320 +/- 9 kg) were fed eight times daily a 46% corn-based concentrate:54% mixed hay diet to meet maintenance energy requirements (3.5 kg of DM/d). In situ production of acetic acid, determined by pulsed-continuous infusion of [1-14C]acetic acid, was 530 mmol/h (CV = 12%). Disappearance from the reticulorumen (i.e., presumed absorption) was 460 mmol/h (CV = 2%) or 87% (CV = 2%) of ruminal production. Variation is described within an operational steady state. Acetic acid concentrations were increased by continuous infusion of unlabeled acetic acid into the reticulorumen. Ruminal disappearance (mmol/h) increased when this simulated production was elevated up to and exceeding in situ rates reported previously (530 to 2,700 mmol/h). These data suggest that two-thirds to three-fourths of ruminal acetate production disappeared across the reticuloruminal wall when concentration was elevated; the complement exited from the rumen with the liquid phase. The reticulorumen has an additional capacity to remove acetic acid; however, it does so with an apparent reduced efficiency at higher production rates. Liquid out-flow may affect the efficiency of reticuloruminal disappearance. Sites distal to the rumen become quantitatively important when ruminal acetic acid concentrations are increased. 相似文献