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21.
Jonathan D. G. Jones David A. Jones Gerard J. Bishop Kate Harrison Bernard J. Carroll Steven R. Scofield 《Transgenic research》1993,2(2):63-78
Cell-autonomous genes have been used to monitor the excision of both endogenous transposons in maize andAntirrhinum, and transposons introduced into transgenic plants. In tobacco andArabidopsis, the streptomycin phosphotransferase (SPT) gene reveals somatic excision of the maize transposonActivator (Ac) as green sectors on a white background in cotyledons of seedlings germinated in the presence of streptomycin. Cotyledons of tomato seedlings germinated on streptomycin-containing medium do not bleach, suggesting that a different assay for transposon excision in tomato is desirable. We have tested the use of the spectinomycin resistance (SPEC) gene (aadA) and a Basta resistance (BAR) gene (phosphinothricin acetyltransferase, or PAT) for monitoring somatic excision ofAc in tobacco and tomato. Both genetic and molecular studies demonstrate that genotypically variegated individuals that carry clones of cells from whichAc orDs have excised from either SPEC or BAR genes, can be phenotypically completely resistant to the corresponding antibiotic. This demonstrates that these genes act non-cell-autonomously, in contrast to the SPT gene in tobacco. Possible reasons for this difference are discussed. 相似文献
22.
The decomposition of the amidinohydrazone-type insecticide Hydramethylnon (HMN) by soil fungi has been investigated. A simple spectrophotometric method was developed for the estimation of HMN in soil and fungal culture media. HMN was found to be degraded in soil with a half life of 14 to 25 days.Degradation of HMN by the lignolytic fungus,Phanerochaete chrysosporium yielded two major breakdown products;p-(trifluoromethyl)-cinnamic acid (TFCA) andp-(trifluoromethyl)-benzoic acid (TFBA). TFCA was converted to TFBA which was subsequently metabolised via themeta-fission pathway. Fluoride release from HMN could not be detected.Abbreviations BzDAc
benzene, dioxane, acetic acid (60: 36: 4)
- DCM
dichloroethane
- DNPH
2,4-dinitro-phenylhydrazine
- HMN
Hydramethylnon
- TDAc
toluene, dioxane, acetic acid (90: 30: 1)
- TFCA
p-(trifluoromethyl)-cinnamic acid
- TFBA
p-(trifluoromethyl)-benzoic acid
- TFP
1,5-bis(trifluoro-p-tolyl)-1,4-pentadien-3-one
- VA
veratryl alcohol 相似文献
23.
Anthony G. Lee Kate A. Dalton Richard C. Duggleby J. Malcolm East Anthony P. Starling 《Bioscience reports》1995,15(5):289-298
Effects of lipid structure on the function of the Ca2+-ATPase of skeletal muscle of sarcoplasmic reticulum are reviewed. Binding of phospholipids to the ATPase shows little specificity. Phosphatidylcholines with short (C14) or long (C24) fatty acyl chains have marked effects on the activity of the ATPase, including a change in the stoichiometry of Ca binding. Low ATPase activity in gel phase lipid follows from low rate of phosphorylation. Phosphatidylinositol 4-phosphate increases ATPase activity by increasing the rate of dephosphorylation of the phosphorylated ATPase. Stimulation is not seen with other anionic phospholipids; phosphatidic acid decreases ATPase activity in a Mg2+-dependent manner.Abbreviations di(C141)PC
dimyristoleoylphosphatidycholine
- di(C160)PC
dipalmitoylphosphatidylcholine
- di(C181)PC
dioleoylphosphatidylcholine
- di(Br2C180)PC
dibromostearoylphosphatidylcholine
- di(C241)PC
dinervonylphosphatidylcholine
- di(C181)PA
dioleoylphosphatidic acid
- di(C181)PE
dioleoylphosphatidylethanolamine
- Ptdlns
phosphatidylinositol
- PtdIns-4P
phos-phatidylinositol 4-phosphate 相似文献
24.
Human medulloblastoma gangliosides 总被引:2,自引:0,他引:2
To establish a model system for the study of ganglioside metabolismof the human brain tumor, medulloblastoma, we have chemicallycharacterized the gangliosides of the Daoy cell line. Thesecells contain a high concentration of gangliosides (143 ±13 nmol LBSA/108 cells). The major species have been structurallyconfirmed to be GM2 (65.9%), GM3 (13.0%), and GDla (10.3%).Isolation of individual gangliosides homogeneous in both carbohydrateand ceramide moieties by reversed-phase HPLC and analysis bynegative-ion fast atom bombardment collisionally activated dissociationtandem mass spectrometry have allowed us to unequivocally characterizeceramide structures. In the case of GM2, 10 major ceramide subspecieswere identified: d18:1-hC16:0, d18:1-C16:0, d18:0-C16:0, d18:1-C18:0,d18:1-C20:0, d18:1-C22:0, d18:2-C24:1, d18: 1-C23:1, d18:1-C24:1,and d18:1-C24:0. Taken together with previous studies, thesefindings in human medullo-blastoma cells support the view thathigh expression and marked heterogeneity of ceramide structureare general characteristics of tumor gangliosides, moleculeswhich are shed by the tumor cells and which are biologicallyactive in vivo. medulloblastoma gangliosides ceramide structure HPLC mass spectrometry 相似文献
25.
26.
H. G. de Vries J. M. Collée W. P. Meeuwsen H. Scheffer L. P. Ten Kate 《Human genetics》1995,95(5):575-576
Number and sex of offspring were determined in a group of 7,841 randomly selected blood donors who were screened for the F508 mutation. We did not find any evidence for differences in number or sex ratio of offspring between F508 carriers and non-carriers. 相似文献
27.
28.
John Leo Abernethy Charles M. Lovett Anton Haddad Jeff D. Felberg 《Bioorganic chemistry》1982,11(3):251-261
Acylated crude papain has been shown to exert stereoselective behavior toward racemic hydrazides devoid of an amino acid residue, namely, (RS)-mandelic and (RS)-atrolactic hydrazides. These hydrazides functioned as nucleophiles to yield N1,N2-diacylhydrazines. Several achiral acylating agents for the enzyme were chosen, including Z-glycine, BOC-glycine, AOC-glycine, and hippuric acid. With the exception of hippuric acid as the acylating agent, the reaction product, in every instance for these achiral hydrazides, consisted of an excess of the (+)-N1,N2-diacylhydrazine. The relative rates of product formation for the mandelic hydrazides were considerably greater than for corresponding reactions with racemic atrolactic hydrazide. When chiral Z-l-alanine was employed to acylate crude papain, the stereoselective action was most pronounced, with the formation of a mixture of diastereoisomers consisting of 73% N1-(Z-l-alanyl)-N2-[(R)-mandelyl]hydrazine. The relative reactivities for the electrophiles was Z-l-alanine ? Z-glycine ? hippuric acid ? AOC-glycine > BOC-glycine. The hydrazides of (R)-, (S)-mandelic, and (RS)-atrolactic acids were prepared by conversion of the corresponding acids to their esters by means of a catalytic dehydrating agent and subsequent treatment with a methanolic solution of hydrazine. 相似文献
29.
John Leo Abernethy Gary F. Kuzmin Charles M. Lovett Jr. William A. Wilson 《Bioorganic chemistry》1980,9(4):440-449
Hydrazides of five N-acylamino acids have been used alone as substrates for papain catalysis to yield N1,N2-diacylhydrazines. With the exception of N-(benzyloxycarbonyl)(Z)-
-alanine hydrazide, they were very effective as both acylating agents of the enzyme and nucleophiles in attacking the enzyme-substrate intermediate. Although Z-
-alanine hydrazide was a minimal acylating agent, it was a satisfactory nucleophile. The most favorable reaction involved Z-
-alanine hydrazide in producing N1,N2-bis(Z-
-alanyl)hydrazine. When Z-
-alanine hydrazide was the substrate, this same chiral diacylhydrazine was formed along with meso N1-(Z-
-alanyl)-N2-(Z-
-alanyl)hydrazine. For the acylation step, the enzyme displayed powerful, essentially stereospecific, bias toward the
enantiomer. Once the thioester intermediate was formed, little preference was detected for attack by the enantiomers as nucleophiles. The most direct procedure for synthesis of substrates was conversion of Z-amino acids to their esters by means of dry HCl in an absolute alcohol. Treatment with hydrazine produced the hydrazides in excellent yield. 相似文献
30.
J E De Vries F H Kornips J Wiegant P M Moerkerk N Senden B Schutte J P Geraedts F T Bosman J Ten Kate 《The journal of histochemistry and cytochemistry》1992,40(7):1053-1058
We describe the combination of hot banding with fluorescence in situ hybridization as a rapid and efficient method to identify integration sites of transfected DNA sequences in chromosomes. As a test system we used SW480 EJ2, a clonal cell line obtained after transfection of SW480 with pSV2neoEJ, a plasmid containing a point-mutated, c-Ha-RAS oncogene. Nick-translated probes were compared with random primed-labeled probes to evaluate their relative efficiency in fluorescence in situ hybridization. The fluorescence signals were quantified in interphase nuclei by confocal scanning laser microscopy. Nick-translated probes were found to yield better results. Hot banding followed by fluorescence in situ hybridization localized the integration site of pSV2neoEJ in SW480 EJ2 at the site of a translocation on a marker chromosome Xp+. The combination of fluorescence in situ hybridization and hot banding can be used to (a) rapidly and efficiently analyze integration sites in large numbers of transfectants, (b) assess the clonality of transfected cell lines, and (c) localize the site of integration of transfected genes in the recipient genome. 相似文献