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961.
Cloned goats (Capra hircus) from adult ear cells 总被引:11,自引:0,他引:11
The average number of available oocytes recovered per ovary collected during the breeding season in dairy goats was 5.5 (1815/330). 66.17% (1201/1815) of oocytes extruded the first polar body after maturation in vitro for 20 h. 75.44% (906/1201) of matured oocytes with membrane evagination around the MII chromosomes were enucleated. Ear skin fibroblast cells were derived from an adult female dining Grey goat (C. hircus). The cells were cryopreserved in liquid nitrogen after passage 2. Thawed cells were further cultured for 3-6 passages and were subjected to serum starvation by 0.5% FBS for 2-10 d, then used as donor cells for nuclear transfer. 98.12% (889/906) of the enucleated oocytes were reconstructed by intracytoplasmic injection of karyoplast. The reconstructed embryos were activated by 5μ mol/L ionomycin for 4.5 min and further activated by culturing with 6-dimethylaminopurine (6-DMAP) for 3 h. After 36 h of culture in mCR1aaBF, 76.69% (645/841) of the cloned embryos cleaved. There were no signifi 相似文献
962.
963.
有机物料对土壤镉形态及其生物有效性的影响 总被引:14,自引:0,他引:14
采用盆栽试验,研究了淹水种稻条件下添加猪粪和泥炭对红壤和潮土中内源和外源Cd形态及其生物有效性的影响。结果表明,土壤中内源Cd在各形态之间的分布比较均匀;添加外源Cd时Cd主要分布于交换态,从分蘖期到成熟期,内源Cd交换态普遍升高,添加外源Cd时交换态普遍降低。有机物料对内源Cd交换态的影响不显著,但当添加外源Cd时则对交换态有显著影响,在不添加外源Cd的条件下,有机物料普遍促进水稻根系对Cd的吸收。在添加外源Cd的条件下,有机物料普遍抑制水稻根系对Cd的吸收,猪粪的抑制效果强于泥炭,水稻根系对Cd与Fe的累积呈显著抑制作用。 相似文献
964.
Solution structure of a yeast ubiquitin-like protein Smt3: the role of structurally less defined sequences in protein-protein recognitions 下载免费PDF全文
Smt3 belongs to a growing family of ubiquitin-related proteins involved in posttranslational protein modification. Independent studies demonstrate an essential function of Smt3 in the regulation of nucleocytoplasmic transport, and suggest a role in cell-cycle regulation. Here we report the high-resolution NMR structure of yeast Smt3 in the complex free form. Our comparison of the Smt3 NMR structure with the Smt3 crystal structure in complex with the C-Terminal Ulp1 protease domain revealed large structural differences in the binding surface, which is also involved in the Smt3-Ubc-9 interaction detected by NMR. The structural differences in the region indicate the important functions of conserved residues in less structurally defined sequences. 相似文献
965.
Kim YJ Kim KP Han SK Munoz NM Zhu X Sano H Leff AR Cho W 《The Journal of biological chemistry》2002,277(39):36479-36488
We reported previously that exogenously added human group V phospholipase A(2) (hVPLA(2)) could elicit leukotriene B(4) (LTB(4)) biosynthesis in human neutrophils (Han, S. K., Kim, K. P., Koduri, R., Bittova, L., Munoz, N. M., Leff, A. R., Wilton, D. C., Gelb, M. H., and Cho, W. (1999) J. Biol. Chem. 274, 11881-11888). To determine the mechanism of the hVPLA(2)-induced LTB(4) biosynthesis in neutrophils, we thoroughly examined the effects of hVPLA(2) and their lipid products on the activity of group IVA cytosolic PLA(2) (cPLA(2)) and LTB(4) biosynthesis under different conditions. As low as 1 nm exogenous hVPLA(2) was able to induce the release of arachidonic acid (AA) and LTB(4). Typically, AA and LTB(4) were released in two phases, which were synchronized with a rise in intracellular calcium concentration ([Ca(2+)](i)) near the perinuclear region and cPLA(2) phosphorylation. A cellular PLA(2) assay showed that hVPLA(2) acted primarily on the outer plasma membrane, liberating fatty acids and lysophosphatidylcholine (lyso-PC), whereas cPLA(2) acted on the perinuclear membrane. Lyso-PC and polyunsaturated fatty acids including AA activated cPLA(2) and 5-lipoxygenase by increasing [Ca(2+)](i) and inducing cPLA(2) phosphorylation, which then led to LTB(4) biosynthesis. The delayed phase was triggered by the binding of secreted LTB(4) to the cell surface LTB(4) receptor, which resulted in a rise in [Ca(2+)](i) and cPLA(2) phosphorylation through the activation of mitogen-activated protein kinase, extracellular signal-regulated kinase 1/2. These results indicate that a main role of exogenous hVPLA(2) in neutrophil activation and LTB(4) biosynthesis is to activate cPLA(2) and 5-lipoxygenase primarily by liberating from the outer plasma membrane lyso-PC that induces [Ca(2+)](i) increase and cPLA(2) phosphorylation and that hVPLA(2)-induced LTB(4) production is augmented by the positive feedback activation of cPLA(2) by LTB(4). 相似文献
966.
Belenkaya TY Han C Standley HJ Lin X Houston DW Heasman J Lin X 《Development (Cambridge, England)》2002,129(17):4089-4101
967.
鸭血清胆碱酯酶的纯化及性质研究 总被引:3,自引:0,他引:3
首次采用新技术双水相萃取方法作为鸭血清胆碱酯酶(EC.3.1.1.8 CHE) 纯化的第一步,后经 DEAE-Sephadex A50,sephadex G200 柱层析,获得电泳纯鸭血清胆碱酯酶,提纯倍数1018倍,酶活力回收43.4%,比活274.9U/mg。鸭血清胆碱酯酶性质研究表明:此酶是糖蛋白和酸性蛋白水解酶,等电点 4.2 左右,最适 pH7.5 左右;对底物碘化硫代丁酰胆碱的 Km=9.8×10-5mol/L;SDS-PAGE 电泳和聚丙烯酰胺梯度电泳表明,鸭血清胆碱酯酶以相同亚基组成的不同聚合体形式存在,亚基分子量 78000,具有完整的酶活性.不同聚合体带电状态相同. 相似文献
968.
Yanhai Yin Shizhong Li Yiming Chen Hongqing Guo Wenzhong Tian Ying Chen Liangcai Li 《Plant Cell, Tissue and Organ Culture》1993,32(1):61-68
Calluses were induced from immature embryos of an indica type rice and finely dispersed cell suspension cultures were initiated from the callus using modified AA medium (S1 medium). The suspension cultures were maintained alternatively (1–2 passages in each medium) in S1 medium and S2 medium, the latter containing KNO3, NH4NO3, proline and glutamine as nitrogen source. Protoplasts of high quality were isolated form suspension cells cultured in S2 medium supplemented with ABA. Embedding the protoplasts in agarose blocks containing NH4NO3-free modified KM8P(PM1) medium and immersing the blocks in NH4NO3-containing modified KM8P(PM3) medium were most effective for obtaining protoplast division and callus formation. The protoplast-derived calluses were precultured in potato extract-aand/or ABA-containing N6(D1, D2 or D3) media and many embryo-like structures were formed. These structures developed into plantlets after being transferred to N6 differentiation (D4) medium. The regenerated plantlets grew into mature plants and beard seeds normally.Abbreviations AA medium
amino acids based medium
- ABA
abscisic acid
- BA
benzyladenine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- DF
division frequency
- IAA
indoleacetic acid
- KIN
kinetin
- NAA
naphthaleneacetic acid
- PE
planting efficiency 相似文献
969.
GP73, a novel Golgi-localized protein upregulated by viral infection 总被引:16,自引:0,他引:16
Kladney RD Bulla GA Guo L Mason AL Tollefson AE Simon DJ Koutoubi Z Fimmel CJ 《Gene》2000,249(1-2):53-65
970.
PCR Detection of Salmonella enterica Serotype Montevideo in and on Raw Tomatoes Using Primers Derived from hilA 总被引:1,自引:0,他引:1 下载免费PDF全文
Salmonellae have been some of the most frequently reported etiological agents in fresh-produce-associated outbreaks of human infections in recent years. PCR assays using four innovative pairs of primers derived from hilA and sirA, positive regulators of Salmonella invasive genes, were developed to identify Salmonella enterica serotype Montevideo on and in tomatoes. Based on examination of 83 Salmonella strains and 22 non-Salmonella strains, we concluded that a pair of hilA primers detects Salmonella specifically. The detection limits of the PCR assay were 101 and 100 CFU/ml after enrichment at 37°C for 6 and 9 h, respectively. When the assay was validated by detecting S. enterica serotype Montevideo in and on artificially inoculated tomatoes, 102 and 101 CFU/g were detected, respectively, after enrichment for 6 h at 37°C. Our results suggest that the hilA-based PCR assay is sensitive and specific, and can be used for rapid detection of Salmonellae in or on fresh produce. 相似文献