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1.
Summary Soluble lead salts and a number of lead-containing minerals catalyze the formation of oligonucleotides from nucleoside 5-phosphorimidazolides. The effectiveness of lead compounds correlates strongly with their solubility. Under optimal conditions we were able to obtain 18% of pentamer and higher oligomers from ImpA. Reactions involving ImpU gave smaller yields.Abbreviations A adenosine - U uridine - Im imidazole - MeIm 1-methyl-imidazole - EDTA ethylenediaminetetraacetic acid - pA adenosine 5-phosphate - pU uridine 5-phosphate - Ap adenosine cyclic 2:3-phosphate - ATP adenosine 5-triphosphate - AppA P1,P2-diadenosine 5-diphosphate - pNp (N = A,U) nucleotide 2(3), 5-diphosphate - ImpA adenosine 5-phosphoreimidazolide - ImpU uridine 5-phosphorimidazolide - A 2pA adenylyl-[25]-adenosine - A 3pA adenylyl-[35]-adenosine - pA 2pA 5-phospho-adenylyl-[25]-adenosine - pA 3pA 5-phospho-adenylyl-[35]-adenosine - pUpU 5-phospho-uridylyl-uridine - pApU 5-phospho-adenylyl-uridine - pUpA 5-phospho-uridylyladenine - (pA)n (n, 2,3,4,) oligoadenylates with 5 terminal phosphate - ImpApA 5-phosphorimidazolide of adenylyl adenosine - (pA) 5+ pentamer and higher oligoadenylates with 5 terminal phosphate - (Ap)nA (n = 2,3,4) oligoadenylates without terminal phosphates In the following we do not specify the nature of the internucleotide linkageIn the following we do not specify the nature of the internucleotide linkage  相似文献   

2.
An anion channel in the plasma membrane of guard cells (GCAC1) provides a regulatory element for the voltage-dependent release of anions during stomatal closure (Keller et al. 1989) as well as excitability (Hedrich et al. 1990). Recognition sites for plant growth hormones on the extracellular surface of GCAC1 further indicate that this channel may also serve as a transduction element in hormone signaling (Marten et al. 1991 a). Stilbene derivatives were used to study the inhibitor-structure channel-function relationship of GCAC1. We have analyzed the activity, voltage-gate and kinetics of this channel as affected by stilbenes. The stilbene derivatives SITS and DNDS caused a shift in activation potential and a decrease in the peak current amplitude. Channel block through the action of DIDS, on the other hand, was not accompanied by a shift in voltage-dependence. Differences in the dose-dependence of the two effects give clues to the presence of channel sites responsible for gate-shifting and block. The ability to inhibit anion currents (Kd) increased in the sequence: SITS (4 µM) < DNDS (0.5 µM) < DIDS (0.2 µM). All inhibitors reversibly blocked the anion channel from the extracellular side. Channel block on the level of single anion-channels is characterized by a reduction of long open-transitions into flickering bursts and a decrease in channel amplitude.Abbreviations DIDS 4,4-Diisothiocyanostilbene-2,2-disulfonic acid - SITS 4-Acetamido-4-isothiocyanostilbene-2,2-disulfonic acid - DNDS 4,4-dinitrostilbene-2,2-disulfonic acid - NPPB 5-Nitro-2-(3-phenylpropylamio)benzoic acid - IAA-94 [(6,7-Dichloro2-cyclopentyl-2,3-dihydro-2-methyl-1-oxo-1H-inden-5y1)oxy] acetic acid - A-9-C Anthracene-9-carboxylic acid - TEA Tetraethylammonium  相似文献   

3.
The 3 ends of chloroplast mRNAs are produced by the processing of longer precursors. The 3 ends of most plastid mRNAs are located at, or several nucleotides downstream of, stem-loop structures, which act as 3-end-processing signals and RNA stability elements. In chloroplasts of the green alga Chlamydomonas reinhardtii, 3-end maturation of atpB mRNA involves endonucleolytic cleavage of the pre-mRNA at an AU-rich site located about 10 nucleotides downstream of the stem-loop structure. This cleavage is followed by exonucleolytic resection to generate the mature 3 end. In order to define critical nucleotides of the endonucleolytic cleavage site, we mutated its sequence. Incubation of synthetic atpB pre-RNAs containing these mutations in a chloroplast protein extract resulted in the accumulation of 3-end-processed products. However, in two cases where the AU-rich sequence of this site was replaced with a GC-rich one, the 3 end of the stable processing product differed from that of the wild-type product. To examine whether these mutations affected atpB mRNA processing or accumulation in vivo, the endogenous 3 UTR was replaced with mutated sequences by biolistic transformation of Chlamydomonas chloroplasts. Analysis of the resulting strains revealed that the accumulation of atpB mRNA was approximately equal to that of wild-type cells, and that a wild-type atpB 3 end was generated. These results imply that Chlamydomonas atpB 3 processing parallels the situation with other endonucleases such as Escherichia coli RNAse E, where specific sequences are required for correct in vitro processing, but in vivo these mutations can be overcome.  相似文献   

4.
The occurrence of adenosine 5-triphosphate-3-diphosphate-synthesizing activity was detected in five strains of actinomycetes; Streptomyces morookaensis, Streptomyces aspergilloides, Streptomyces hachijoensis, Actinomyces violascens and Streptoverticillium septatum, out of 825 strains of actinomycetes, bacteria, fungi and imperfecti. Purine nucleotide pyrophosphotransferase were extracellularly excreted associating with the cell growth, and were purified partially or to apparent homogeniety from the culture filtrate. The enzymes are a monomeric protein with a molecular weight of 18000–26000 and synthesize adenosine, guanosine and inosine 5-phosphate (mono, di or tri)-3-diphosphate such as pApp, ppApp, pppApp, pGpp, ppGpp, pppGpp and pppIpp by transferring a pyrophosphoryl group from the 5-position of ATP, dATP and pppApp to the 3-position of purine nucleotides in the presence of a divalent cation and in alkaline state.Abbreviations pppApp adenosine 5-triphosphate 3-diphosphate - ppApp adenosine 5-diphosphate 3-diphosphate - pApp adenosine 5-monophosphate 3-diphosphate - pppGpp guanosine 5-triphosphate 3-diphosphate  相似文献   

5.
A comparison of the effects of ionic stress and an uncoupler on long-term fluorescence transients (the Kautsky effect) in the green alga Dunaliella tertiolecta indicated that the large quenching induced by ionic stress was caused by a pH gradient across the thylakoid membrane. This possiblity was given support by the increase in the slow phase of 3-(3,4-dichlorophenyl)-1,1-dimethylurea-induced fluorescence relaxation in algae subjected to ionic stress. Low-temperature fluorescence emission spectra indicated that salt stress enhanced photosystem-I emission in the dark, and a comparison of simultaneous emissions at 695 and 720 nm at room temperature indicated a further increase in photosystem-I emission during the fluorescence transients. Taken together with the decrease in the fast phase of 3-(3,4-dichlorophenyl)-1,1-dimethylurea-induced fluorescence relaxation in stressed algae, our results indicate that ionic stress stimulates cyclic electron flow, and that non-cyclic flow is inhibited. The effect of sucrose-induced osmotic stress was similar to, but less marked than, the effects of NaCl and KCl; the effect of decreasing the external salinity was small.Abbreviations DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - FCCP carbonylcyanide p-trifluoromethoxyphenylhydrazone - PSI, II photosystem I, II  相似文献   

6.
The cleavage of adenosine-5-monophosphate (5-AMP) and guanosine-5-monophosphate (5-GMP) by Ce4+ and lanthanide complex of 2-carboxyethylgermanium sesquioxide (Ge-132) in acidic and near neutral conditions was investigated by NMR , HPLC and measuring the liberated inorganic phosphate at 37°C and 50°C. The results showed that 5-GMP and 5-AMP was converted to guanine (G), 5-monophosphate (depurination of 5-GMP), ribose (depurination and dephosphorylation of 5-GMP), phosphate and adenine (A), 5-monophosphate (depurination of 5-AMP), ribose (depurination and dephosphorylation of 5-AMP), phosphate respectively by Ce4+. In presence of lanthanide complexes, 5-GMP and 5-AMP were converted to guanosine (Guo) and phosphate and adenosine (Ado) and phosphate respectively. The mechanism of cleaving 5-GMP and 5-AMP is hydrolytic scission  相似文献   

7.
Summary The carotenoid pigments of the myxobacterium Sorangium compositum were analyzed by chromatographical and chemical techniques and by visible, infra red, and mass spectroscopy. Besides -carotene, neurosporene, torulene, lycopene, and 1,2-dihydro-1-hydroxy--carotene, four new carotenoid glycosides were found. These pigments were identified as 1,2-dihydro-1-hydroxy-torulene glucoside ester (I), 1,2-dihydro-3,1-dihydroxy-torulene glucoside ester (III), 1,2-dihydro-1-hydroxy-torulene rhamnoside (II), and 1,2-dihydro-3,1-dihydroxytorulene rhamnoside (IV).Fifth communication on the carotenoids of myxobacteria. Fourth communication see Arch. Mikrobiol. 76, 364–380 (1971).  相似文献   

8.
A monoclonal antibody generated against synthetic peptides patterned on amino acids 542–555 of human band 3, designated 1F4, specifically immunostainedPlasmodium falciparum-infected erythrocytes and inhibited the cytoadherence ofP. falciparum-infected erythrocytes to C32 amelanotic melanoma cells. 1F4 did not recognize intact band 3 protein on immunoblots, however it was reactive towards proteolytic fragments of band 3.The binding region of another murine monoclonal antibody previously reported to recognize the membrane spanning domain of human band 3, designated B6, was found to also recognize residues 542–555, however its properties differed from 1F4. Mab B6 recognized both infected and uninfected red cells, and reacted only with intact band 3 on immunoblots. Mab B6 was without effect on cytoadherence.These results demonstrate that monoclonal antibodies reactive against a common peptide sequence may bind to different conformations of the peptide sequence and suggest that the adherent competency ofP. falciparum-infected erythrocytes may result from a change in the surface topography of human band 3 protein.Abbreviations ELISA Enzyme-Linked Immunosorbent Assay - KLH Keyhole Limpet Hemocyanin - PBS Phosphate Buffered Saline - Mab Monoclonal Antibody - PMSF Phenylmethyl sulfonyl fluoride - i.p. intraperitoneum - TBS Tris Buffered Saline - H2DIDS dihydro 4,4-diisothocyanostilbene-2,2-disulfonic acid - DIDS 4,4-diisothiocyanostilbene-2,2-disulfonic acid  相似文献   

9.
The origin of Q-independent derivatives of phage lambda   总被引:13,自引:0,他引:13  
Summary qsr (Q-independent) phages are characterised by the replacement of the region of the genome that contains Q, S, R, and the late gene promoter, PR, with host-derived DNA that codes for functions analogous to those deleted. Restriction endonuclease analysis and DNA/DNA hybridisation methods have been used to show that p4 and qin A 3, two such Q-independent phages, are the product of recombination between and a defective lambdoid prophage (the qsr prophage) located at an as yet unidentified site in the E. coli K 12 chromosome. The qsr prophage is distinct from the defective lambdoid prophage Rac (Kaiser and Murray 1979). In the E. coli K 12 strain AB1157 from which qsr phages cannot be generated, the qsr prophage has suffered an internal deletion. That the qsr prophage appears not to carry a full complement of essential late genes suggests one explanation for its apparently defective nature.  相似文献   

10.
Summary Suspensions of endocrine pancreas cells were prepared by shaking collagenase-isolated rat islets of Langerhans in calcium-free buffer. When incubated with 1.0 mM substrate at pH 7.4, the cells split,P i from 5-AMP at a rate of 87 nmol/h per g DNA, and from-glycerophosphate at a rate of 25 nmol/h per g DNAK m for 5 AMP was about 54 M. Adenosine or theophylline inhibited the 5-AMP hydrolysis. Homogenization of the cells increased the activity toward 5-AMP by 23% and that toward-glycerophosphate by 115%. Injecting rats with cortisone had no effect on the 5-AMP hydrolysis by whole cells but significantly increased the activity in cell homogenates; the intracellular activity toward 5-AMP was more than doubled by the cortisone treatment. Staining whole islet cells for 5-AMP-splitting activity resulted in a demarcation of the cell periphery in control rats. Cells from cortisone-treated rats showed heavier deposits of reaction product, and their cell periphery did not stand out as clearly. It is suggested that 5-nucleotidase is largely an ectoenzyme in normal rat islet cells. The cells also contain an as yet unidentified intracellular phosphatase that seems to be solely responsible for the increased hydrolysis of 5-AMP in cortisone-treated rats.  相似文献   

11.
Maria Mulisch 《Protoplasma》1988,143(2-3):170-175
Summary Different fixation techniques were employed to obtain satisfactory fixation of the endoplasm ofStentor coeruleus for ultrastructural investigations. The nuclei ofS. coeruleus are surrounded by a flattened fenestrated cisterna. The space between the nuclear envelope and the cisterna (= perinuclear space) is continuous with the cytoplasm via channels. The envelopes of both, micronucleus and macronucleus, are connected to the fenestrated cisterna by filamentous material. This organization accounts for the close association between micronucleus and macronucleus inStentor coeruleus. The fenestrated cisterna is compared to similar structures occurring in other organisms, and its possible function is discussed.Abbreviations fC fenestrated cisterna - FV food vacuole - km km fibers - MaNu macronucleus - MiNu micronucleus - My myonome - NE nuclear envelope - PC perinuclear cisterna - PfC pore of fenestrated cisterna - PS perinuclear cytoplasmic space - EGTA ethyleneglycol-bis-(-aminoethylether)-N,N,N,N-tetraacetic acid) - GA glutaraldehyde - HEPES N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid - PEG polyethylene glycol - PIPES piperazine-N,N-bis[2-ethanesulfonic acid] - PTA phosphotungstic acid  相似文献   

12.
The carotenoid pigments of a Rhizobium strain isolated from Lotononis bainesii were found to be diglucosyl-4,4-diapocarotene-4,4-dioate and glucosyl-4,4-diapocarotene-4-oate-4-oic acid.5th publication in the series Carotenoids of Rhizobia [4th publication: Helv. chim. Acta 62: 2551–2557 (1979)]  相似文献   

13.
The sugar conformation of a DNA decamer was studied with proton-proton 3J coupling constants. Two samples, one comprising stereospecifically labeled 2-R-2H for all residues and the other 2-S-2H, were prepared by the method of Kawashima et al. [J. Org. Chem. (1995) 60, 6980–6986; Nucleosides Nucleotides (1995) 14, 333–336], the deuterium labeling being highly stereospecific 99% for all 2-2H, 98% for 2-2H of A, C, and T, and 93% for 2-2H of G). The 3J values of all H1-H2 and H1-H2 pairs, and several H2-H3 and H2-H3 pairs were determined by line fitting of 1D spectra with 0.1–0.2 Hz precision. The observed J coupling constants were explained by the rigid sugar conformation model, and the sugar conformations were found to be between C3-exo and C2-endo with m values of 26° to 44°, except for the second and 3 terminal residues C2 and C10. For the C2 and C10 residues, the lower fraction of S-type conformation was estimated from JH1H2 and JH1H2 values. For C10, the N–S two-site jump model or Gaussian distribution of the torsion angle model could explain the observed J values, and 68% S-type conformation or C1-exo conformation with 27° distribution was obtained, respectively. The differences between these two motional models are discussed based on a simple simulation of J-coupling constants.  相似文献   

14.
Plausible prebiotic conditions for the phosphorylation of nucleosides by inorganic phosphate were reported by Lohrmann and Orgel in 1971. This reaction was carried out on heated dry films and promoted by urea. The major products formed were nucleoside-2:3 cyclicPs; 5-NMPs and other derivatives were also formed. Minor modifications of the Lohrmann and Orgel system have resulted in the preferential formation of 5-NMPs. In this modified system a 2-fold preference for phosphorylation of the 5-OH group over the 3(2)-OH group was observed and the formation of other derivatives was minimized. The small amounts of bis compounds that were formed in this system could be quantitatively removed by selective binding to the mineral hydroxylapatite at moderate ionic strengths. It was also discovered that under hydrolytic conditions there was a 3:1 preference for removal of phosphates attached to the 3-OH group over the 5-OH group. A recycling procedure for obtaining additional 5-NMPs from bis compounds and 3-NMPs is proposed.  相似文献   

15.
Binding of the fluorophore 1,1-bis(4-anili-no) naphthalene-5,5-disulfonic acid (bis-ANS) to influenza virus A/PR 8/34 is strongly enhanced at low pH. Binding is accompanied by a significant increase in fluorescence intensity. The binding and the fluorescence increase are associated with the low-pH induced conformational change of the viral spike protein, hemagglutinin, exposing hydrophobic binding sites. The data indicate that in addition to the hydrophobic N-terminus of HA2 other hydrophobic sequences of the HA ectodomain become accessible to bis-ANS at low pH. It is shown that the time course of the fluorescence increase of bis-ANS at low pH is determined by the conformational change of HA. The application of this assay for continuously monitoring the kinetics of the structural alteration in HA is discussed and its relevance for elucidating the temporal relationship between the conformational change of HA and virus-membrane fusion is outlined.Abbreviations HA hemagglutinin - BHA bromelain-solubilized ectodomain of HA - N-HA2 N-terminus of the HA2 subunit - PBS phosphate buffered saline - bis-ANS (1,1-bis(4-anilino)naphthalene-5,5-disulfonic acid) - R18 octadecylrhodamine B chloride - FDQ fluorescence dequenching - RBC red blood cell Correspondence to: A. Herrmann  相似文献   

16.
M. Grün  G. Franz 《Planta》1981,152(6):562-564
Biosynthetic studies with cell-free extracts from Aloe arborescens Mill. demonstrate the transfer of the glucose moiety from UDP-glucose to aloe emodin anthrone, forming the C-glycosidic linkage in the anthracene derivative aloin. The pH-dependence and the specificity of UDP-glucose and aloe emodin anthrone for the biosynthesis of the C-glycosidic bond in aloin are shown.Abbreviations ADP-Glc adenosine-5-diphosphate glucose - AEA aloe emodin anthrone (1,8-dihydroxy-3-(hydroxymethyl)-9(10 H)-anthracenone) - CoASAc acetyl coenzyme A - GDP-Glc guanosine-5-diphosphate glucose - Glc glucose - Glc-1-P glucose-1-phosphate - HPLC high performance liquid chromatography - TLC thin layer chromatography - UDP-Gal uridine-5-diphosphate galactose - UDP-Glc uridine-5-diphosphate glucose  相似文献   

17.
4-Methylumbelliferyl 6-O-benzyl--d-lactoside (6Bn-MU-Lac) and some related compounds were synthesizedvia different selective reactions including phase-transfer glycosylation. Their suitability as substrates for a fluorometric assay of ceramide glycanase (CGase) was evaluated. Among others, the 6Bn-MU-Lac, which is resistant to exogalactosidase, was found to be a suitable substrate for routine assay of the CGase activity. For American leech CGase, theK m value is 0.232 mM at pH 5. Abbreviations: CGase, ceramide glycanase; Gal, galactose; Glc, Glucose; Lac, lactose; MU, 4-methylumbelliferone; MU-Lac, 4-methylumbelliferyl -d-lactoside; bBn-Lac, 6-O-benzyl-lactose; 6Bn-MU-Lac, 4-methylumbelliferyl 6-Obenzyl--d-lactoside; 46Bd-MU-Lac, 4-methylumbelliferyl 4,6-O-benzylidene--d-lactoside; MU-Cel, 4-methylumbellifery -d-cellobioside; 46Bd-MU-Cel, 4-methylumbelliferyl 4,6-O-benzylidene--d-cellobioside; TLC, thin layer chromatography;1H-NMR, proton nuclear magnetic resonance; GSL, glycosphingolipids; CSA, 10-camphorsulfonic acid. See Scheme 1 for chemical structures.  相似文献   

18.
Summary Physarum polycephalum microplasmodia exposed to 1.6×10–5 M cytochalasin A evidenced intracellular cytoplasmic condensation, slow contraction, and eventual breaks at discrete surface areas, within one hour. Other cytochalasins tested (CB or CD) did not substitute for CA. CA effects on plasmodia were not abolished by immediate washing or media replacement. In nutrient medium, CA plus ATP (375 M) produced within minutes herniation (blebbing) and plasmodial disruption. The order of addition of reagents was important; ATP added simultaneously with or prior to CA stimulated the phenomenon, whereas initial addition of CA resulted in no such dynamic response. Several other nucleotides (e.g., AMP, cAMP) could substitute for ATP; however, such changes were not observed with 5-adenylylimidodiphosphate. Blebbing was not abolished in the presence of 2,4-dinitrophenol. In minimal medium, it was best stimulated by simultaneous addition of Ca++ and Mg++. Preincubation of CA with L-cysteine or with -mercaptoethanol negates its individual or nucleotide-combined effects. Yet, 10–5 M ethacrynic acid, a sulfhydryl-reactive liposoluble drug, in the presence of ATP does not mimic the blebbing response. These observed effects, which take place at or near the plasmodial surface, presumably reflect acceleration of normal contractile processes inPhysarum. Abbreviations CA cytochalasin A - CB cytochalasin B - CD cytochalasin D - AMP adenosine 5-monophosphate - ADP adenosine 5-diphosphate - ATP adenosine 5-triphosphate - di-butyryl-cAMP di-butyryl-cyclic adenosine 35-monophosphate - di-butyrylcGMP di-butyryl-cyclic guanosine 35-monophasphate. This work was supported by a grant (AI-11902) from the U.S. Public Health Service.  相似文献   

19.
Summary Anaerobic conditions obtained by flooding the soil caused reductive dechlorination of p,p-DDT and its conversion to p,p-DDD was enhanced under water-logged conditions creating or favouring anaerobiosis. The DDT showed recalcitrance in the soil kept at 15% moisture.More o,p-DDT was lost from the flooded soil. Similar amounts of p,p-DDE were detected in all of the three levels of technical DDT treatments and the concentrations were not significantly different under both aerobic and anaerobic conditions.  相似文献   

20.
Summary The self-condensation of 2(3)-O-glycyl esters of adenosine, adenosine-5-(O-methylphosphate) and P1, P2-diadenosine-5-pyrophosphate in 6.2 mM solutions at pH 8.0 and -5°C in the presence of 12.5 mM poly(U) yields approximately 3 times as much diketopiperazine as reactions without poly(U). As the concentration of 2(3)-O-(glycyl)-P1, P2-diadenosine-5-pyrophosphate is decreased from 6.2 mM to 1.5 mM the yield of diketopiperazine in the presence of poly(U) decreases slightly from 6.6% to 5.2%, whereas, in the absence of poly(U) the yield of diketopiperazine decreases substantially from 2.4% to 0.75%. The enhanced yield of diketopiperazine that is attributed to the template action of poly(U) is temperature dependent and is observed only at temperatures below 10°C (5°C to -5°C) for 6.2 mM 2(3)-O-(glycyl)-adenosine-5-(O-methylphosphate) and below 23°C (15°C to -5°C) for 6.2 mM 2(3)-O-(glycyl)-P1, P2-diadenosine-5-pyrophosphate. The absence of a template effect at high temperatures is attributed to the melting of the organized helices. The hydrolysis half-lives at pH 8.0 and -5°C of 2(3)-O-(glycyl)-adenosine, 2(3)-O-(glycyl)-adenosine-5-(O-methylphosphate), 2(3)-O-(glycyl)-P1, P2-diadenosine-5-pyrophosphate, and 5-O-(glycyl)-adenosine in the presence of poly(U) are substantially larger than their half-lives in the absence of poly(U). The condensation of 2(3)-O-(glycyl)-adenosine yields 5% of 5-O-(glycyl)-adenosine in the presence of poly(U) compared to 0.7% in the absence of poly(U).Abbreviations DKP diketopiperazine - (gly)2 glycylglycine - (gly)3 glycylglycylglycine - AppA-gly 2(3)-O-(glycyl)-P1, P2-diadenosine-5-pyrophosphate - MepA-gly 2(3)-O-(glycyl)-adenosine-5-(O-methylphosphate) - Ado-2(3)-gly 2(3)-O-(glycyl)-adenosine - Ado-5-gly 5-O-(glycyl)-adenosine - Boc-gly N-tert-butyloxycarbonylglycine - AppA P1, P2-diadenosine-5-pyrophosphate - MepA adenosine-5-(O-methylphosphate) - AppA-Boc-gly 2(3)-O-(Boc-glycyl)-P1, P2-diadenosine-5-pyrophosphate - Ado-5-Boc-gly 5-O-(Boc-glycyl)-adenosine - Ado-2(3)-Boc-gly 2(3)-O-(Boc-glycyl)-adenosine  相似文献   

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