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111.
利用PCR方法对单纯疱疹病毒Ⅱ型糖蛋白D(HSV-2gD)基因进行了修饰,在其5'端删去约500bp的非编码区,仅保留ATG上游7个bp。将修饰后的HSV-2gD基因插入到带有痘苗病毒天坛株TK基因区段的痘苗表达质粒pJSA1175,置于痘苗病毒P7.5k早/晚期启动子控制下。将此重组质粒用脂质体Lipofectin方法转染已受野型TK ̄+痘苗病毒天坛株感染的TK ̄-143细胞,通过同源重组机制和标志基因LacZ产物的蓝斑显色作用,以及BudR试剂对TK表型的选择压力,筛选出整合有HSV-2gD基因的重组痘苗病毒。Southem杂交表明,HSV-2gD基因已正确地插入痘苗病毒TK基因区内;间接免疫荧光检测显示,HSV-2gD蛋白已得到有效表达,且主要分布于细胞膜。重组病毒免疫家兔可产生明显的抗HSV-2gD中和抗体。用重组病毒免疫小鼠,3周后可使94%(17/18)的小鼠对抗HSV-2的致死量攻击,表明重组病毒具有明显的免疫保护作用。 相似文献
112.
THELASERTOMOGRAPHICALMETHODUSINGMINIMUMOFPROJECTIONFORBIOLOGICALOBJECTSTRUCTURESTUDYYuriN.Kulchin;OlegB.Vitrik;OlegV.Kirichei... 相似文献
113.
Guo-Ling Nan Adelheid R. Kuehnle 《In vitro cellular & developmental biology. Plant》1995,31(3):131-136
Summary Five parameters were examined for their effect on transformation ofDendrobium tissues by microprojectile bombardment. The superpromoter in pBI426 produced at least 1.5 times as many transient transformants
as the single cauliflower mosaic virus 35S promoter in pBI121 (37 to 69% vs. 0 to 44%) with dark and frequent GUS (β-glucuronidase) staining. Tissue, genotype, and type of microparticle significantly affected transient GUS activity. Higher
expression was seen in protocormlike bodies and in hybrid UH44 compared to etiolated shoots and protocorms and to hybrids
M61 and K1329-39. Microparticles of 1.6-μm Bio-Rad gold were more effective than 1.0-μm ASI gold. Transient GUS activity did
not differ among protocormlike bodies bombarded using helium propellant pressures of 650, 900, or 1100 psi. Transgenic plants
were recovered fromDendrobium UH800 protocormlike bodies bombarded with pBI426-coated, 1.1-μm tungsten particles using an early-model gunpowder-driven
apparatus with an estimated stable transformation rate of 11.7%. One transgenic plant ofDendrobium UH44 was recovered from etiolated shoot explants bombarded with pBI121-coated, 1.1-μm tungsten particles using the Dupont
PDS-1000 with a stable transformation rate of 0.17%. Positive selection results showed 100 to 200 mg·liter−1 kanamycin to be appropriate for regeneration of transgenic plants from protocormlike bodies, protocorms, and etiolated shoot
explants over a 3- to 9.5-mo. period. 相似文献
114.
Josiane Girardie Adrien Girardie Alain Van Dorsselaer Odile Sorokine Serge Geoffre Michel Hospital Gilles Precigoux 《Journal of peptide science》1995,1(5):311-318
An original insect neurohormone of 65 residues was synthesized by the solid-phase methodology using t-Boc strategy and Boc-Val-PAM–resin. The purification, conducted by several steps of liquid chromatography having mass, polarity or charge as separative criteria, yielded the product with the correct molecular weight of 6922 Da determined by mass spectrometry. The synthetic peptide had both the same affinity for the antinative neurohormone serum and the same biological activity as the native neurohormone. 相似文献
115.
Ralph Rapley 《Molecular biotechnology》1995,3(2):139-154
The exquisite specificity of monoclonal antibodies (MAb) has long provided the potential for creating new reagents for the
in vivo delivery of therapeutic drugs or toxins to defined cellular target sites or improved methods of diagnosis. However,
many difficulties associated with their production, affinity, specificity, and use in vivo have largely confined their application
to research or in vitro diagnostics. This situation is beginning to change with the recent developments in the applied molecular
techniques that allow the engineering of the genes that encode antibodies rather than the manipulation of the intact antibodies
themselves. Techniques, such as the polymerase chain reaction, have provided essential methods with which to generate and
modify the genetic constituents of antibodies, allow their conjugation to toxins or drugs, provide ways of humanizing murine
antibodies, and allow discrete modular antigen binding components to be produced. More recent developments of in vitro expression
systems and powerful phage surface display technologies will without doubt play a major role in future antibody engineering
and in the successful development of new diagnostic and therapeutic antibody-based reagents. 相似文献
116.
Functional response of Neoseiulus barkeri Hughes on two-spotted spider mite (Acari: Tetranychidae) 总被引:1,自引:0,他引:1
Generalist predatory mites are the common phytoseiid fauna in many agroecosystems, but little attention has been paid to their potential as biological control agents. In this study, we determined the functional responses of adult females of the generalist predator Neoseiulus barkeri Hughes on eggs, larvae, and adults of the two-spotted spider mite, Tetranychus urticae Koch, in the laboratory. Predation experiments were conducted on pepper leaf discs over a 24 h period at 25±1°C, 70–80% RH and 16L:8D photoperiod. Prey densities ranged 5 to 80 eggs, or 5 to 40 larvae, or 1 to 8 female adults of T. urticae per disc. The predation rate of N. barkeri adult females on T. urticae eggs was the same as on its larvae, but the predation rate on adult females was much lower. The role of generalist predatory mites in integrated and biological control of greenhouse pests was discussed. 相似文献
117.
M J WILSON D M GLEN S K GEORGE J D PEARCE C W WILTSHIRE 《The Annals of applied biology》1994,125(2):377-390
A field experiment on winter wheat in autumn 1991 investigated the effect of the rhabditid nematode, Phasmarhabditis hermaphrodita, applied to soil at five dose rates (108 - 1010 infective larvae ha-1) immediately after seed sowing, on slug populations and damage to seeds and seedlings. The nematode was compared with methiocarb pellets broadcast at recommended field rate immediately after drilling and no molluscicide treatment. Slug damage to wheat seeds and seedlings was assessed 6 and 13 wk after drilling. Seedling survival increased and slug grazing damage to seedlings declined linearly with increasing log nematode dose. These two measures of slug damage were combined to give an index of undamaged plant equivalents, which also increased linearly with increasing log nematode dose. ANOVA showed that, after 6 wk, there were significantly more undamaged plant equivalents on plots treated with the two highest nematode doses (3 × 109 and 1 × 1010 ha-1) than on untreated plots, but the number of undamaged plant equivalents on methiocarb-treated plots was not significantly greater than that on untreated plots. Slug populations were assessed by refuge trapping and soil sampling. Deroceras reticulatum was the commonest of several species of slugs recorded. During the first 4 wk after sowing, significantly more slugs were found under refuge traps on plots treated with certain doses of P. hermaphrodita than under traps on untreated plots and more showed signs of nematode infection than expected from the prevalence of infection in slugs from soil samples, suggesting that the presence of P. hermaphrodita altered slug behaviour. Application of P. hermaphrodita had no significant impact on numbers or biomass of slugs in soil during a 27 wk period after treatment, except after 5 wk when slug numbers were inversely related to log nematode dose. However, by this time, numbers in soil samples from untreated plots had declined to levels similar to those in plots treated with the highest dose of nematodes. During the first 5 wk after treatment, c. 20% of slugs in soil samples from untreated plots showed symptoms of nematode infection. It is suggested that this represented the background level of infection in the experimental field rather than spread of infection from treated plots. The apparent lack of impact of P. hermaphrodita on slug numbers and biomass in soil suggests that its efficacy in protecting wheat from slug damage was through inhibition of feeding by infected slugs. 相似文献
118.
Y. Hailemichael J. W. Smith R. N. Wiedenmann 《Entomologia Experimentalis et Applicata》1994,71(2):155-166
Xanthopimpla stemmator (Thunberg)(Hymenoptera: Ichneumonidae), a solitary endoparasite of pupae of Old World lepidopteran stalkborers, was recently
imported into Texas as a candidate for biological control of New World stalkborers. Information on host acceptability, host
suitability and cues responsible for host finding were necessary to gain an insight into parasite/host interactions, because
of the absence of a coevolutionary history.Xanthopimpla stemmator females were exposed to laboratory-reared one-to six-day-oldDiatraea saccharalis (F.) pupae. An average of 62% of host pupae were accepted and all ages of pupae were equally acceptable. Host suitability
decreased with host age. One- to five-day-old host pupae averaged 31–37% suitability, whereas only 19% of 6-day-old pupae
were suitable. Successful parasitization, defined as the product of the proportion accepted and the proportion suitable, decreased
from 22–23% for 1-, 2- and 3-day-old pupae to 13% for 6-day-old pupae. Sex ratio (female:male) of the parasite progeny increased
with host age. Females comprised 47% of total parasite progeny of 1-day-old and 84% of 6-day-old pupae. The increase in percent
females was a result of a similar number of females in all age classes, coupled with a decrease in the number of males from
older hosts.Xanthopimpla stemmator superparasitized 61% of acceptedD. saccharalis pupae in the laboratory. On dissection, 73% of host pupae with multiple probe wounds were found to contain parasite eggs
or larvae; these hosts contained up to 10 eggs or 7 first-instar larvae. Increased numbers of probes by the parasites were
associated with an increase in successful parasitization. Host seeking activity inX. stemmator was stimulated by the presence of larval frass, host odor and movement of host pupae. Results suggest thatX. stemmator is a good candidate for biological control ofD. saccharalis and possibly other factitious stalkborer hosts. 相似文献
119.
Changes in basin geomorphology after implementation of the Oosterschelde estuary project 总被引:8,自引:8,他引:0
The completion in 1986/87 of an open storm-surge barrier in the inlet and of secondary dams in the landward parts of the Oosterschelde tidal basin (SW Netherlands) has had and will continue to have a significant impact on geomorphological developments. An analysis of historic data, and of recent detailed bathymetric and morphodynamic process data, indicates that former trends have reversed. At present the Oosterschelde is a sedimentation basin with a degrading intertidal area and silting up of channels. The continuing reduction in intertidal area, the decreasing geomorphological gradients, the increasing fine sediment content of channel deposits, combined with a general reduction in hydrodynamics, imply significant ecological effects. 相似文献
120.
Pires MJ Martel P Baptista A Petersen SB Willson RC Cabral JM 《Biotechnology and bioengineering》1994,44(7):773-780
The mechanism of extraction of rat cytochrome b(5) from water into a sodium dioctylsulfosuccinate (AOT) micellar organic phase was studied using protein engineering of surface charged residues. The extraction behavior of native cytochrome b(5) and modified proteins with substitutions of the type glutamic acid --> lysine at positions 44 (E44K), 56 (E56K), and 92 (E92K), was studied as a function of pH. The results indicate that an important mechanism of extraction is an electrostatic interaction of this protein with the negatively charged surfactant. We demonstrate that it is possible to improve extraction by engineering the protein surface charge, increasing the driving force responsible for the protein transfer to the micellar phase. (c) 1994 John Wiley & Sons, Inc. 相似文献