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61.
本文提取人骨骼肌α辅肌动蛋白(α-actinin)是综合了文献报导有关提取兔肌α-actinin的和提取鸡胗α-actinin的方法,稍加修改而确定的。用Hasselbach-Schneider缓冲液提取骨骼肌中的肌球蛋白后,将残余物经硼酸-缓冲液提取、匀浆及高速离心去掉肌动蛋白和肌原纤维的其它成份,上清加硫酸铵至30%,35%饱合度所得的沉淀用220mmol/LTris-乙酸溶解、透析、离心后经DE-52柱层析可得电泳纯。α-actinin。将人骨骼肌α-actinin纯化制品免疫了三只大耳白纯种家兔,两个多月后,三只兔子都产生免抗人骨骼肌α-actinin的特异抗血清,用双向免疫扩散法和酶联免疫吸附试验(ELISA)测定,产生的抗体效价较高,用双扩散法测定效价为1:32,用ELISA测定,用比率法判断结果,效价最高者为1:100,000左右,经免疫电镜观察结果证实,上述抗血清可以满足进一步实验要求。 相似文献
62.
The Ih and lh
i alleles have been shown previously to reduce the level of endogenous gibberellin A1 (GA1) in shoots of pea (Pisum sativum L.), resulting in a dwarf phenotype compared with the wild type, cv. Torsdag (Lh). In addition, plants homozygous for the lh
i allele have reduced seed yield compared with Lh (tall, wild type) and lh (dwarf) plants. In this paper we show that the lh
i mutation is expressed in developing seeds and pods. Comparison of GA levels in young shoots and developing seeds of genotypes lh and lh
i demonstrates that the relative severity of the two mutations varies in different tissues. Homozygous h
i seeds have reduced GA levels, weigh less, and are less likely to develop to maturity when compared with Lh seeds. However, fertilization of lh
i plants with Lh pollen increases seed GA levels, seed weight and seed survival, indicating that an increase in seed GA levels due to the presence of the Lh allele can restore normal seed growth. Pods developing on self-pollinated lh
i plants are shorter than pods on Lh (wild type) plants, although this may be an indirect effect of the increased seed abortion of lh
i plants. Based on these results we suggest that endogenous GAs play an important role in the development of seeds of P. sativum L.Abbreviations GA(n)
gibberellin An
We wish to thank Katherine McPherson, Peter Newman, Leigh Johnson and Peter Bobbi for technical assistance, Professor L. Mander (ANU, Canberra) and Professor B.O. Phinney (UCLA, USA) for labelled GA standards, and the Australian Research Council for financial support. 相似文献
63.
H. M. Jahns 《Plant Systematics and Evolution》1993,187(1-4):145-174
Fragments of several lichens were grown in the laboratory and developmental steps of growth and regeneration could be described. Different degrees of determination could be observed. The stages of podetial development inCladonia are easily distinguished and are reached after comparably short time. As they occur highly synchronized the results encourage further morphogenetic experiments.Dedicated to Prof. DrO. L. Lange on the occasion of his retirement as a tribute to his outstanding qualities as both a scientist and as a teacher and humane colleague. 相似文献
64.
65.
Efforts to leverage clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) for targeted genomic modifications in mammalian cells are limited by low efficiencies and heterogeneous outcomes. To aid method optimization, we developed an all-in-one reporter system, including a novel superfolder orange fluorescent protein (sfOrange), to simultaneously quantify gene disruption, site-specific integration (SSI), and random integration (RI). SSI strategies that utilize different donor plasmid formats and Cas9 nuclease variants were evaluated for targeting accuracy and efficiency in Chinese hamster ovary cells. Double-cut and double-nick donor formats significantly improved targeting accuracy by 2.3–8.3-fold and 19–22-fold, respectively, compared to standard circular donors. Notably, Cas9-mediated donor linearization was associated with increased RI events, whereas donor nicking minimized RI without sacrificing SSI efficiency and avoided low-fidelity outcomes. A screen of 10 molecules that modulate the major mammalian DNA repair pathways identified two inhibitors that further enhance targeting accuracy and efficiency to achieve SSI in 25% of transfected cells without selection. The optimized methods integrated transgene expression cassettes with 96% efficiency at a single locus and with 53%–55% efficiency at two loci simultaneously in selected clones. The CRISPR-based tools and methods developed here could inform the use of CRISPR/Cas9 in mammalian cell lines, accelerate mammalian cell line engineering, and support advanced recombinant protein production applications. 相似文献
66.
R. M. WARWICK 《Zoological Journal of the Linnean Society》1989,96(3):229-241
The meiobenthos are regarded as the first metazoans to appear on earth This paper considers some aspects of their role in the marine ecosystem over evolutionary time. It is speculatively suggested that they have had a profound influence on many structural and functional attributes of the marine ecosystem as we see it today, both in the benthic and pelagic realms. Certain features such as size spectra, the life-history characteristics of the macrobenthos, and the energetic balance between ecological compartments of the marine ecosystem are considered to result from evolution away from negative interactions with the meiobenthos. 相似文献
67.
Abstract: Post-embryonic development is controlled by two types of meristems: apical and lateral. There has been considerable progress recently in understanding the function of root and shoot apical meristems at the molecular level. Knowledge of analogous processes in the lateral, or secondary, meristems, i.e. the vascular cambium or cork cambium, is, however, rudimentary. This is despite the fact that much of the diversity in the plant kingdom is based on the differential functions of these meristems, emphasizing the importance of lateral meristems in the development of different plant forms. The vascular cambium is particularly important for woody plants, but it also plays an important role during the development of various herbaceous species, such as Arabidopsis thaliana. In this review, we focus on the two basic functions of cambial activity: cell proliferation and pattern formation. 相似文献
68.
C. J. M. Stevens G. te Kronnie J. Samallo H. Schipper H. W. J. Stroband 《Development genes and evolution》1996,205(7-8):460-467
A subtractive-hybridization technique, combined with differential screenings and subsequent whole mount in situ hybridization (ISH) reactions, was used to isolate novel cDNA clones representing developmentally-regulated genes of carp. Small-scale differential screenings of an oocyte and a segmentation-stage cDNA library using oocyte-specific and segmentation stage-specific enriched probes, yielded 75 positive clones. ISH screening showed that 65% (15) of the oocyte-stage clones and 50% (26) of the segmentation-stage clones were indeed stage-specific. Partial sequence analysis suggests that approximately 65% of the 41 stage-specific clones represent novel genes. In addition, an Otxl clone was isolated. Two novel clones and the Otxl clone are of special interest for developmental studies. The clones represent genes that are locally expressed during embryonic development. The expression patterns of Otxl and one of the novel clones suggest functions in specification of the anterior-posterior axis. The three clones provide molecular markers for the study of gastrulation and the patterning of the a-p axis in teleosts. 相似文献
69.
Increased intracellular calcium concentration ([Ca2+]i) is required for smooth muscle contraction. In tracheal and other tonic smooth muscles, contraction and elevated [Ca2+]i are maintained as long as an agonist is present. To evaluate the physiological role of steady-state increases in Ca2+ on tension maintenance, [Ca2+]i was elevated using ionomycin, a Ca2+ ionophore or charybdotoxin, a large-conductance calcium-activated potassium channel (KCa) blocker prior to or during exposure of tracheal smooth muscle strips to Ach (10–9 to 10–4
M). Ionomycin (5 µM) in resting muscles induced increases in [Ca2+]i to 500±230 nM and small increases in force of 2.6±2.3 N/cm2. This tension is only 10% of the maximal tension induced by ACh. Charybdotoxin had no effect on [Ca2+]i or tension in resting muscle. After pretreatment of muscle with ionomycin, the concentration-response relationship for ACh-induced changes in tension shifted to the left (EC50=0.07±0.05 µM ionomycin; 0.17±0.07 µM, control, p<0.05). When applied to the muscles during steady-state responses to submaximal concentrations of ACh, both ionomycin and charybdotoxin induced further increases in tension. The same magnitude increase in tension occurs after ionomycin and charybdotoxin treatment, even though the increase in [Ca2+]i induced by charybdotoxin is much smaller than that induced by ionomycin. We conclude that the resting muscle is much less sensitive to elevation of [Ca2+]i when compared to muscles stimulated with ACh. Steady-state [Ca2+]i limits tension development induced by submaximal concentrations of ACh. The activity of KCa moderates the response of the muscle to ACh at concentrations less than 1 µM. 相似文献
70.
M. J. Kushmerick 《Journal of bioenergetics and biomembranes》1995,27(6):555-569
Muscular activity converts chemical energy into useful work and metabolism restores muscle to its original state. This essay explores the organization and interactions of the regulatory system(s) which allow this energy balance to occur. The term energy balance is used in a biochemical rather than a thermodynamic sense—concerned not with deductions from the physical principles of thermodynamics, but rather with those enzymatic processes which nature evolved and which operate at remarkably fixed stoichiometry. Energy balance is a statement of conservation of energy put into biochemical observables.31P NMR spectroscopy is one of the most useful techniques for investigating these questions quantitatively under physiological conditionsin vivo. The author (1) describes the rules or principles of biochemical energy balance; (2) discusses sample results from human muscle to demonstrate its use in studying this class of questions; (3) presents a simple model of integrated cellular respiration to demonstrate its sufficiency to account for the main observations. 相似文献