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51.
苏云金芽孢杆菌δ-内毒素基因穿梭质粒的构建 总被引:1,自引:0,他引:1
在农业生产中长期使用化学农药已对环境和生态平衡造成一定破坏作用,同时有不少害虫也逐渐产生抗药性从而引起某些害虫的大流行,给农业生产带来巨大损失。应用苏云金杆菌杀虫蛋白基因(Bt基因)可构建具有抗虫作用的抗虫工程菌,这样通过拌种或植物叶面喷雾可达到快速、经济、有效的防治虫害的目的。国际上抗虫工程菌研究应用很快,如美国将BI基因转入到一种正常情况下定居在植物组织中的棒杆菌,将这种工程菌拌玉米种子,这样随植物生长该菌在植物体内大量繁殖,当玉米螟在茎和叶取食时,即因食用表达苏云金杆菌毒蛋白的工程菌而死亡。 田颖川等已克隆了苏云金芽孢杆菌内毒素基因CryIA(b)和CryIA(c)。本文将Bt基因CryIA(c)插入到大肠-枯草穿梭载体pBE-2中构建成Bt毒蛋白基因穿梭质粒pAMY,利用电穿孔法转人大肠杆菌DH5a,枯草芽孢杆菌B.subtilis BR151,IA511,野生型蜡状芽孢杆菌B.cereusa-47,短芽孢杆菌B.brevis A-5和枯草芽孢杆菌90-8,获得了具有较高杀虫活性的工程菌克隆。 相似文献
52.
Cloning of the macrolide antibiotic biosynthesis gene acyA, which encodes 3-O-acyltransferase, from Streptomyces thermotolerans and its use for direct fermentative production of a hybrid macrolide antibiotic. 总被引:2,自引:0,他引:2
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A Arisawa N Kawamura K Takeda H Tsunekawa K Okamura R Okamoto 《Applied microbiology》1994,60(7):2657-2660
A gene encoding the macrolide modification enzyme 3-O-acyltransferase (acyA) was cloned by chromosome walking onto the carbomycin biosynthetic region in Streptomyces thermotolerans TH475, with the 3' region of the gene encoding the macrolide modification enzyme 4"-O-acyltransferase (acyB1) as a probe. A shortened fragment (1.8 kb) containing acyA was subcloned with pIJ350. A high-level tylosin producer, Streptomyces fradiae MBBF, transformed with the plasmid could produce a hybrid macrolide, 3-O-acetyltylosin, most efficiently. 相似文献
53.
Atomic composition of the hydrophobic and hydrophilic membrane sides of self-assembled SC3p hydrophobin.
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H A Wsten O M de Vries H C van der Mei H J Busscher J G Wessels 《Journal of bacteriology》1994,176(22):7085-7086
The hydrophobin SC3p of Schizophyllum commune self-assembles into a 10-nm-thick amphipathic membrane at hydrophilic-hydrophobic interfaces. X-ray photoelectron spectroscopy of the hydrophobic membrane side of SC3p, assembled in vitro, showed an atomic composition similar to the calculated composition of SC3p when glycosylation was taken into account. The atomic composition measured at the hydrophilic membrane side deviated from that at the hydrophobic side and indicated the presence of a lower number of peptide bonds. High levels of S and N were detected only on mycelia carrying hydrophobic aerial hyphae, as expected with assembled SC3p present at the surface of these hyphae. 相似文献
54.
Abscisic Acid-Induced Membrane Potential Changes in Barley Aleurone Protoplasts: a Possible Relevance for the Regulation of rab Gene Expression 总被引:2,自引:0,他引:2
Heimovaara-Dijkstra Sjoukje; van Duijn Bert; Libbenga Kees R.; Heidekamp Freek; Wang Mei 《Plant & cell physiology》1994,35(5):743-750
The effect of ABA on the membrane potential of barley (Hordeumvulgare cv. Himalaya) aleurone protoplasts was studied by measuringthe distribution of the lipophilic cation tetraphenylphosphonium(TPP+). The resting membrane potential (Em) according to ourmeasurements with TPP+ is about 53 mV and is in agreementwith membrane potential values as measured with intracellularmicroelectrodes (about 55 mV). The TPP+-measurementscould demonstrate a clear dependence of the resting Em on theexternal pH (pHe). Stimulation of the protoplasts with ABA induced a transienthyperpolarization of the membrane to 62 mV as measuredwith TPP+. The hyperpolarization was ABA-concentration dependent. Inhibition of the H+-ATPases with the specific proton pump inhibitorsdiethylstilbestrol (DES) or Micanozole effectively preventedhyperpolarization. This indicates that the hyperpolarizationis consistent with the activation of plasma membrane H+-ATPases.The K+-inward rectifier inhibitor BaCl2 was able to prolongthe hyperpolarization. This result suggests that the hyperpolarizationcauses the opening of K+-channels. The ABA-induced proton-pump activation may be involved in ABA-inducedgene-expression, as DES was able to inhibit this gene expression.BaCl2 did only show a slight inhibitory effect on ABA-inducedgene-expression. (Received January 4, 1994; Accepted April 12, 1994) 相似文献
55.
A procedure for the mass propagation of multiple shoots of Stevia rebaudiana is described. Isolated shoot primordia were used as the inoculum to obtain clusters of shoot primordia. Such clusters were grown in a 500 liter bioreactor to obtain shoots. A total of 64.6 Kg of shoots were propagated from 460 g of the inoculated shoot primordia. These shoots were easily acclimatized in soil. 相似文献
56.
Inorganic cofactors (Mn, Ca2+ and Cl-) are essential for oxidation of H2O to O2 by Photosystem II. The Mn reductants NH2OH and its N-methyl derivatives have been employed as probes to further examine the interactions between these species and Mn at the active site of H2O oxidation. Results of these studies show that the size of a hydroxylamine derivative regulates its ability to inactivate O2 evolution activity, and that this size-dependent inhibition behavior arises from the protein structure of Photosystem II. A set of anions (Cl-, F- and SO4
2-) is able to slow NH2OH and CH3NHOH inactivation of intact Photosystem II membranes by exerting a stabilizing influence on the extrinsic 23 and 17 kDa polypeptides. In contrast to this non-specific anion effect, only Cl- is capable of attenuating CH3NHOH and (CH3)2NOH inhibition in salt-washed preparations lacking the 23 and 17 kDa polypeptides. However, Cl- fails to protect against NH2OH inhibition in salt-washed membranes. These results indicate that the attack by NH2OH and its N-methyl derivatives on Mn occurs at different sites in the O2-evolving complex. The small reductant NH2OH acts at a Cl--insensitive site whereas the inhibitions by CH3NHOH and (CH3)2NOH involve a site that is Cl- sensitive. These findings are consistent with earlier studies showing that the size of primary amines controls the Cl- sensitivity of their binding to Mn in the O2-evolving complex.Abbreviation MES
4-morpholinoethanesulfonic acid
- PS II
Photosystem II 相似文献
57.
人肝刺激因子对大鼠实验性慢性肝损伤的保护作用 总被引:3,自引:0,他引:3
从健康孕妇水囊引产4─6个月龄的胎儿取肝,采用LaBrecque方法提取人肝刺激因子(hHSS)。经3H-胸腺嘧啶核苷参入肝DNA法测定其生物活性。表明此hHSS可刺激肝细胞DNA合成。采用皮下注射CCl4和饮用10%乙醇来制备慢性肝损伤动物模型,观察了hHSS的保护肝脏作用。结果表明:hHSS可使CCl4-乙醇所致慢性肝损伤大鼠的死亡率、血清谷丙转氨酶水平、肝组织中羟脯氨酸含量的升高以及肝组织中丙二醛的含量降低。肝组织切片表明:hHSS能减轻肝组织的损伤程度,促进肝细胞再生,并能明显防止肝纤维化的形成和发展。可见,hHSS对CCl4-乙醇所致的慢性肝损伤大鼠有明显的保护作用,其机制可能与促进肝细胞再生及抑制肝细胞膜的脂质过氧化有关。 相似文献
58.
单纯疱疹病毒(HSV)Ⅰ型及Ⅱ型之间有很多共同抗原,能引起血清学交叉反应,鉴别诊断比较困难。本实验利用重组DNA技术,将部分HSV-2DNA的PstI片段克隆到载体质粒PSK中,并筛选出两个重组质粒(P和P)只与HSV-2反应,与HSV-1不反应,这两个重组质粒中所含的HSV-2DNA片段大小分别是3.1和4.3kb,另外,还筛选了一个重组质粒(PHSV2-1,含5.8kbHSV-2DNA片段)与HSV-1和HSV-2均反应。将4.3kb的片段用光生物素标记后作为探针检测了159份人阴道拭子,其中23份样品呈阳性反应,其余均为阴性,从23份阳性样品中挑选12价涂片用间接荧光抗体法检测也都呈阳性反应,随机挑选的几份杂交反应阴性样品在间接荧光试验中也是阴性。本实验制备的HSV通用及HSV-2型特异性探针将比常规的血清学方法诊断和鉴别HSV-1和HSV-2感染更为可靠。 相似文献
59.
用Northern blot方法对二乙基亚硝胺所诱发的大鼠肝癌中内源性蛋白酶抑制因子α_2-巨球蛋白(α_2-M)、非特异性免疫抑制剂α_1-酸性糖蛋白(α_1-AGP)及雄性激素正调控的α-2u球蛋白(α-2u)三种分泌性蛋白基因表达情况进行了分析。结果表明在大部分(14/16)肝癌样品中α_2-M RNA水平显著降低;而α_1-AGP RNA水平显著高于正常对照水平;α-2u RNA水平明显下降,但在某些雄性大鼠肝癌样品中该基因却有一定程度的表达。这些结果说明,一些肿瘤宿主血浆中α_2-M水平的显著下降及α_1-AGP水平的明显升高分别是由于基因表达活性的下降及升高所致。α-2u基因表达的异常提示,在癌变过程中机体的内分泌功能发生了某些变化。 相似文献
60.
Ziliang Li Weijie Xu Luyao Kang Yakov Kuzyakov Leiyi Chen Mei He Futing Liu Dianye Zhang Wei Zhou Xuning Liu Yuanhe Yang 《Global Change Biology》2023,29(22):6367-6382
Mineralization of dissolved organic matter (DOM) in thermokarst lakes plays a non-negligible role in the permafrost carbon (C) cycle, but remains poorly understood due to its complex interactions with external C and nutrient inputs (i.e., aquatic priming and nutrient effects). Based on large-scale lake sampling and laboratory incubations, in combination with 13C-stable-isotope labeling, optical spectroscopy, and high-throughput sequencing, we examined large-scale patterns and dominant drivers of priming and nutrient effects of DOM biodegradation across 30 thermokarst lakes along a 1100-km transect on the Tibetan Plateau. We observed that labile C and phosphorus (P) rather than nitrogen (N) inputs stimulated DOM biodegradation, with the priming and P effects being 172% and 451% over unamended control, respectively. We also detected significant interactive effects of labile C and nutrient supply on DOM biodegradation, with the combined labile C and nutrient additions inducing stronger microbial mineralization than C or nutrient treatment alone, illustrating that microbial activity in alpine thermokarst lakes is co-limited by both C and nutrients. We further found that the aquatic priming was mainly driven by DOM quality, with the priming intensity increasing with DOM recalcitrance, reflecting the limitation of external C as energy sources for microbial activity. Greater priming intensity was also associated with higher community-level ribosomal RNA gene operon (rrn) copy number and bacterial diversity as well as increased background soluble reactive P concentration. In contrast, the P effect decreased with DOM recalcitrance as well as with background soluble reactive P and ammonium concentrations, revealing the declining importance of P availability in mediating DOM biodegradation with enhanced C limitation but reduced nutrient limitation. Overall, the stimulation of external C and P inputs on DOM biodegradation in thermokarst lakes would amplify C-climate feedback in this alpine permafrost region. 相似文献