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1.
Mammalian brain tubulin is an heterodimer; both and exist in 6–7 isotypic forms which differ in their amino acid sequences. By the use of isotype-specific monoclonal antibodies, we have previously shown that we can purify the
II,
III, anda
IV tubulin dimers from bovine brain. We have also observed that these isotypes differ in their distributionin vivo and their polymerization and drug-binding propertiesin vitro. We have now explored the question of whether the isotypically purified dimers differ in their overall conformation using as probes compounds of theN,N-polymethylenebis (iodoacetamide) series which are known to form discrete intrachain cross-links in-tubulin. These compounds have the structure ICH2CONH(CH2)
n
NHCOCH2I. One of these cross-links, designated
s, is between cys12 and either cys201 or cys211. The other, designated
*, is between cys239 and cys354. The
* cross-link forms in
II and
IV but not in
III; this is not surprising in view of the fact that
III has serine at position 239 instead of cysteine. However,
III is also unable to form the
s cross-link, although it appears to have all three cysteines which may be involved in the cross-link. This suggests that at least one of the sulfhydryls involved in the cross-link may be inaccessible in
III. Although both
II and
IV can form the
s cross-link, the dependence on cross-linker chain length is different.
II forms
s with derivatives in whichn=2, 4, 5, 6, and 7 but not with those in whichn=3 or 10. In contrast,
IV forms
s with derivatives in whichn=2, 3, 4, 5, 6, 7, and 10. These results imply that the
s sulfhydryls are slightly more accessible in
IV and are therefore less dependent on the conformation of the cross-linker to react with it. It appears, therefore, that the
II,
III, and
IV dimers each have unique conformations. This may help to explain the different assembly and drug-binding properties of these dimers. 相似文献
2.
Laura M. M. Ottoboni Adilson Leite Jose A. Yunes Maria Luiza P. N. Targon Gonçalo A. de Souza Filho Paulo Arruda 《Plant molecular biology》1993,21(5):765-778
Several genomic and cDNA clones encoding the 22 kDa-like -coixin, the -prolamin of Coix seeds, were isolated and sequenced. Three contiguous 22 kDa-like -coixin genes designated -3A, -3B and -3C were found in the 15 kb -3 genomic clone. The -3A and -3C genes presented in-frame stop codons at position +652. The two genes with truncated ORFs are flanking the -3B gene, suggesting that the three -coixin genes may have arisen by tandem duplication and that the stop codon was introduced before the duplication.Comparison of the deduced amino acid sequences of -coixin clones with the published sequences of 22 kDa -zein and 22 kDa-like -kafirin revealed a highly conserved protein structure. The protein consists of an N-terminus, containing the signal peptide, followed by ten highly conserved tandem repeats of 15–20 amino acids flanked by polyglutamines, and a short C-terminus. The difference between the 22 kDa-like -prolamins and the 19 kDa -zein lies in the fact that the 19 kDa protein is exactly one repeat motif shorter than the 22 kDa proteins.Several putative regulatory sequences common to the zein and kafirin genes were identified within both the 5 and 3 flanking regions of -3B. Nucleotide sequences that match the consensus TATA, CATC and the ca. –300 prolamin box are present at conserved positions in -3B relative to zein and kafirin genes. Two putative Opaque-2 boxes are present in -3B that occupies approximately the same positions as those identified for the 22 kDa -zein and -kafirin genes. Southern hybridization, using a fragment of a maize Opaque-2 cDNA clone as a probe, confirmed the presence of Opaque-2 homologous sequences in the Coix and sorghum genomes.The overall results suggest that the structural and regulatory genes involved in the expression of the 22 kDa-like -prolamin genes of Coix, sorghum and maize, originated from a common ancestor, and that variations were introduced in the structural and regulatory sequences after species separation. 相似文献
3.
Summary Five subunits (-, -, -, - and -subunits) of the six -and -subunits) in the F1 portion (F1ATPase) of sweet potato (Ipomoea batatas) mitochondrial adenosine triphosphatase were isolated by an electrophoretic method. The - and -subunits were not distinguishable immunologically but showed completely different tryptic peptide maps, indicating that they were different molecular species. In vitro protein synthesis with isolated sweet potato root mitochondria produced only the -subunit when analyzed with anti-sweet potato F1ATPase antibody reacting with all the subunits except the -subunit. Sweet potato root poly(A)+RNA directed the synthesis of six polypeptides which were immunoprecipitated by the antibody: two of them immunologically related to the -subunit and the others to the - and -subunits. We conclude that the -subunit of the F1ATPase is synthesized only in the mitochondria and the -, - and -subunits are in the cytoplasm. 相似文献
4.
The white rot basidiomycete Phanerochaete chrysosporium metabolized 4-ethoxy-3-methoxyphenyl-glycerol--guaiacyl ether (V) in low nitrogen, stationary cultures under which conditions the ligninolytic enzyme system is expressed. 4-Ethoxy-3-methoxyphenylglycerol XIII, guaicol and 4-ethoxy-3-methoxybenzyl alcohol (II) were isolated as metabolic products. Exogenously added XIII was rapidly converted to 4-ethoxy-3-methoxybenzyl alcohol indicating that it is an intermediate in the metabolism of V. P. chrysosporium also metabolized 1-(4-ethoxy-3-methoxyphenyl)-2-(2-methoxyphenoxy)-3-hydroxypropane VI. The degradation pathway for this dimer also included initial -ether cleavage and -hydroxylation of the diol product 1-(4-ethoxy-3-methoxyphenyl) 2,3 dihydroxypropane (XI) to yield the triol XIII which was cleaved at the , bond to yield 4-ethoxy-3-methoxybenzyl alcohol. Finally P. chrysosporium also cleaved the dimer 1-(4-ethoxy-3-methoxyphenyl)-2-(2-methoxyphenoxy)-1-hydroxypropane (VIII) at the -ether linkage yielding 1-(4-ethoxy-3-methoxyphenyl) 1,2 dihydroxypropane (IX) which was subsequently cleaved at the , bond to yield II. All of the results indicate that oxidative -ether cleavage is an important initial reaction in the metabolism of -aryl ether lignin substructure dimeric compounds. Metabolities were identified after comparison with chemically synthesized standards by gas liquid chromatography-mass spectrometry.Abbreviations GLC
Gas liquid chromatography
- TMSi
trimethylsilyl
- TLC
thin layer chromatography 相似文献
5.
The purified isopropylmalate synthase of Alcaligenes eutrophus H 16 reacted with the following -keto acids and acyl-coenzyme A derivatives (in the sequence of decreasing affinities): -ketoisovalerate, -keto-n-valerate, -ketobutyrate and pyruvate; acetyl-CoA, propionyl-CoA, butyryl-CoA. malonyl-CoA, valeryl-CoA, and crotonyl-CoA. -Ketoisocaproate, however, is a strong inhibitor of the enzyme. All reactions catalyzed by isopropylmalate synthase were inhibited to the same extent by the endproduct l-leucine. the substrate saturation curves of -ketoisovalerate or other -keto acids and of acetyl-coenzyme A or other acyl-CoA derivatives had intermediary plateau regions; the Hill coefficient alternated between n
H
-values higher and lower than 1.0, indicating changes from positive to negative and from negative to positive cooperativity for the substrates. The products, isopropylmalate and free coenzyme A, showed competitive inhibition patterns against both substrates (-ketoisovalerate and acetyl-CoA). Free coenzyme A (1 M) inactivated the enzyme irreversibly. The 3-phosphate of coenzyme A and the free carboxyl group of -ketoisovalerate were involved in optimal binding of these substrates, but 3-dephospho-acetyl-coenzyme A and the methylester of -ketoisovalerate were also converted by this enzyme. A CH3–CH2-grouping of the -keto acids seemed to be necessary for binding this substrate.Abbreviations Used CoA
Coenzyme A
- Tris
Tris(hydroxymethyl)aminomethane hydrochloride
- DTNB
5,5-dithiobis-(2-nitrobenzoic acid)
- IPM
-Isopropylmalate
- KIV
-Ketoisovalerate
Prepared from doctoral thesis of the University of Göttingen 1973 相似文献
6.
Jun-Ichiro Murayama Hitoshi Manabe Kayoko Fukuda Hideo Utsumi Akira Hamada 《Glycoconjugate journal》1989,6(4):499-510
Sialic acids and the majorO-glycosidic oligosaccharide of glycophorin MK from monkey (Japanese monkey,Macaca fuscata) erythrocyte membranes were characterized.N-Glycolylneuraminic acid (neu5Gc) was found as the major sialic acid, which was confirmed by gas-liquid chromatography-mass spectrometry as the trimethylsilyl methyl ester. ThreeO-glycosidic oligosaccharide units were obtained from a tryptic glycopeptide that contained all of the carbohydrate units in glycophorin MK by mild alkaline borohydride/borotritide treatment. Carbohydrate analyses of the oligosaccharides revealed that they were composed of Neu5Gc, galactose andN-acetylgalactosaminitol in the molar ratios of 111 (trisaccharide), 211 (tetrasaccharide) and 111 (pentasaccharide). The content of oligosaccharide units was estimated to be 1125 for penta-, tetra- and trisaccharide, respectively, based on the yields, the molecular weight, and the number of oligosaccharide attachment sites in the amino-acid sequence. The tetrasaccharide was the major oligosaccharide and its structure was proposed to be Neu5Gc2-3Gal1-3[Neu5Gc2-6]GalNAcol. 相似文献
7.
Guiyun Xu Takashi Suzuki Yasuhiro Maejima Tohru Mizoguchi Mamoru Tsuchiya Makoto Kiso Akira Hasegawa Yasuo Suzuki 《Glycoconjugate journal》1995,12(2):156-161
The sialidase of swine influenza A viruses of N1 and N2 subtypes, isolated from 1930 to 1992, was studied for substrate specificity with ganglio-series, lacto-series type II and GM3 gangliosides containing Neu5Ac2-3Gal, Neu5Gc2-3Gal and Neu5Ac2-6Gal linkages. All viral sialidases tested showed that the activity for hydrolysing substrates with Neu5Ac2-3Gal was higher than the activities with Neu5Gc2-3Gal and Neu5Ac2-6Gal linkages. When GM1b, GM3 and sialylparagloboside were used as substrates, the earliest strain (A/Wisconsin/15/30 H1N1, isolated in 1930) showed the activity ratio of Neu5Ac2-6Gal to Neu5Ac2-3Gal to be 0.13:0.2, and the ratio Neu5Gc2-3Gal/Neu5Ac2-3Gal to be 0.19:0.37, while those strains isolated from 1978 to 1992 exhibited ratios of 0.29:0.58 for Neu5Ac2-6Gal/Neu5Ac2-3Gal and 0.51:0.76 for Neu5Gc2-3Gal/Neu5Ac2-3Gal. The above results indicate that the substrate specificities of sialidases from swine influenza A viruses towards sialyl linkages and the molecular species of sialic acid are related to the year of isolation, i.e. strains isolated after 1978 exhibited higher activity towards Neu5Ac2-6Gal and Neu5Gc2-3Gal linkages when compared with strains isolated in an earlier year, 1930.Abbreviation Neu5Ac
5-N-acetylneuraminic acid
- Neu5Gc
5-N-glycolyneuraminic acid
- Gal
d-galactose
- Glc
d-glucose
- Cer
Ceramide
- II3(Neu5Ac)Lac
Neu5Ac2-3Gal1-4Glc
- GM3(Neu5Ac2-3Gal)
Neu5Ac2-3Gal1-4Glc1-Cer
- GM3(Neu5Gc2-3Gal)
Neu5Gc2-3Gal1-4Glc1-Cer
- GM1b(Neu5Ac2-3Gal)
Neu5Ac2-3Gal1-3GalNac1-4Gal1-4Glc1-Cer
- GMlb(Neu5Gc2-3Gal)
Neu5Gc2-3Gal1-3GalNAc1-4Gal1-4Glc1-Cer
- IV3(Neu5Ac)nLc4Cer
Neu5Ac2-3Gal1-3GlcNAc1-4Gal1-4Glc1-Cer
- IV3(Neu5Gc)nLc4Cer
Neu5Gc2-3Gal1-3GlcNAc1-4Gal1-4Glc1-Cer
- IV6(Neu5Ac)nLc4Cer
Neu5Ac2-6Gal1-3GlcNAc1-4Gal1-4Glc1-Cer
- TDC
taurodeoxycholate. 相似文献
8.
The overlapping fragments of the chromosomal DNA from black widow spider Latrodectus mactans carrying genes for high-molecular-mass protein neurotoxins, - and -latroinsectotoxins (-LIT and -LIT) and -latrotoxin (-LTX), were PCR-amplified and cloned. Restriction analysis of the PCR products showed that the distribution and sizes of the restriction fragments coincided with those deduced from the earlier sequencing of cDNAs of the corresponding genes. It thus followed that the -LIT and -LIT genes are intronless. Along with our data on the structure of the -latrocrustotoxin (-LCT), this implies that the lack of introns is a common feature of the black widow spider genes encoding high molecular mass neurotoxins. 相似文献
9.
P. E. Cheshev E. A. Khatuntseva Yu. E. Tsvetkov A. S. Shashkov N. E. Nifantiev 《Russian Journal of Bioorganic Chemistry》2004,30(1):60-70
4-O-Glycosylation of 2-azidoethyl 2,3,6-tri-O-benzyl-4-O-(2,3-di-O-benzyl-6-O-benzoyl--D-galactopyranosyl)--D-glucopyranoside with a disaccharide donor, 4-trichloroacetamidophenyl 4,6-di-O-acetyl-2-deoxy-3-O-(2,3,4,6-tetra-O-acetyl--D-galactopyranosyl)-1-thio-2-trichloroacetamido--D-galactopyranoside, in dichloromethane in the presence of N-iodosuccinimide and trifluoromethanesulfonic acid resulted in a tetrasaccharide, 2-azidoethyl (2,3,4,6-tetra-O-acetyl--D-galactopyranosyl)-(1 3)-(4,6-di-O-acetyl-2-deoxy-2-trichloroacetamido--D-galactopyranosyl)-(1 4)-(2,3-di-O-benzyl-6-O-benzoyl--D-galactopyranosyl)-(1 4)-2,3,6-tri-O-benzyl--D-glucopyranoside, in 69% yield. The complete removal of O-protecting groups in the tetrasaccharide, the replacement of N-trichloroacetyl by N-acetyl group, and the reduction of the aglycone azide group to amine led to the target aminoethyl glycoside of -D-Gal-(1 3)--D-GalNAc-(1 4)--D-Gal-(1 4)--D-Glc-OCH2CH2NH2 containing the oligosaccharide chain of asialo-GM1 ganglioside in 72% overall yield. Selective 3-O-glycosylation of 2-azidoethyl 2,3,6-tri-O-benzyl-4-O-(2,6-di-O-benzyl--D-galactopyranosyl)--D-glucopyranoside with thioglycoside methyl (ethyl 5-acetamido-4,7,8,9-tetra-O-acetyl-3,5-dideoxy-2-thio-D-glycero--D-galacto-2-nonulopyranosyl)oate in acetonitrile in the presence of N-iodosuccinimide and trifluoromethanesulfonic acid afforded 2-azidoethyl [methyl (5-acetamido-4,7,8,9-tetra-O-acetyl-3,5-dideoxy-D-glycero--D-galacto-2-nonulopyranosyl)oate]-(2 3)-(2,6-di-O-benzyl--D-galactopyranosyl)-(1 4)-2,3,6-tri-O-benzyl--D-glucopyranoside, the selectively protected derivative of the oligosaccharide chain of GM3 ganglioside, in 79% yield. Its 4-O-glycosylation with a disaccharide glycosyl donor, (4-trichloroacetophenyl-4,6-di-O-acetyl-2-deoxy-3-O-(2,3,4,6-tetra-O-acetyl--D-galactopyranosyl) 1-thio-2-trichloroacetamido--D-galactopyranoside in dichloromethane in the presence of N-iodosuccinimide and trifluoromethanesulfonic acid gave 2-azidoethyl (2,3,4,6-tetra-O-acetyl--D-galactopyranosyl)-(1 3)-(4,6-di-O-acetyl-2-deoxy-2-trichloroacetamido--D-galactopyranosyl)-(1 4)-{[methyl (5-acetamido-4,7,8,9-tetra-O-acetyl-3,5-dideoxy-D-glycero--D-galacto-2-nonulopyranosyl)onate]-(2 3)}-(2,6-di-O-benzyl--D-galactopyranosyl)-(1 4)-2,3,6-tri-O-benzyl--D-glucopyranoside in 85% yield. The resulting pentasaccharide was O-deprotected, its N-trichloroacetyl group was replaced by N-acetyl group, and the aglycone azide group was reduced to afford in 85% overall yield aminoethyl glycoside of -D-Gal-(1 3)--D-GalNAc-(1 4)-[-D-Neu5Ac-(2 3)]--D-Gal-(1 4)--D-Glc-OCH2CH2NH2 containing the oligosaccharide chain of GM1 ganglioside. 相似文献
10.
The extracellular surface of the-chain ofTorpedo california acetylcholine receptor (AChR) was mapped for regions that are accessible to binding with antibodies against a panel of synthetic overlapping peptides which encompassed the entire extracellular parts of the chain. The binding of the antipeptide antibodies to membrane-bound AChR (mbAChR) and to isolated, soluble AChR. was determined. The specificity of each antiserum was narrowed down by determining the extent of its cross-reaction with the two adjacent peptides that overlap the immunizing peptide. With mbAChR, high antibody reactivity was obtained with antisera against peptides1–16,89–104,158–174,262–276, and388–408. Lower, but significant, levels of reactivity were obtained with antibodies against peptides67–82,78–93,100–115, and111–126. On the other hand, free AChR bound high levels of antibodies against peptides34–49,78–93,134–150,170–186, and194–210. It also bound moderate levels of antibodies against peptides262–276 and388–408. Low, yet significant, levels of binding were exhibited by antibodies against peptides45–60,111–126, and122–138. These binding studies, which enabled a comparison of the accessible regions in mbAChR and free AChR, revealed that the receptor undergoes considerable changes in conformation upon removal from the cell membrane. The exposed regions found here are discussed in relation to the functional sites of AChR (i.e., the acetylcholine binding site, the regions that are recognized by anti-AChR antibodies, T-cells and autoimmune responses and the regions that bind short and long neurotoxins).Abbreviations used AChR
acetylcholine receptor
-
mbAChR
membrane-bound AChR
- BSA
bovine serum albumin
- BTX
-bungarotoxin
- EAMG
experimental autoimmune myasthenia gravis
- MG
myasthenia gravis
- PBS
0.15 MNaCl in 0.01 M sodium phosphate buffer, pH 7.2. 相似文献
11.
Mycolic acid patterns of some species of Mycobacterium 总被引:5,自引:0,他引:5
David E. Minnikin S. Megan Minnikin James H. Parlett Michael Goodfellow Mogens Magnusson 《Archives of microbiology》1984,139(2-3):225-231
Representative strains of some species of Mycobacterium were degraded by both acid and alkaline methanolysis. Two-dimensional thin-layer chromatography was used to determine the patterns of mycolic acids and other long-chain components in these methanolysates. Patterns composed of -, methoxy- and ketomycolates were found in Mycobacterium asiaticum, Mycobacterium bovix, Mycobacterium gastri, Mycobacterium gordonae, Mycobacterium kansasii, Mycobacterium marinum and Mycobacterium tuberculosis; a representative of Mycobacterium thermoresistibile also contained lower molecular weight -mycolates in addition to these three acids. In representatives or Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium nonchromogenicum, Mycobacterium novum, Mycobacterium paratuberculosis, Mycobacterium scrofulaceum, Mycobacterium terrae, Mycobacterium xenopi, and Mycobacterium sp. MNC 165 - and ketomycolates were accompanied by -carboxymycolates and 2-eicosanol and homologous alcohols which are derived from wax-ester mycolates. Mycobacterium fortuitum and Mycobacterium giae contained - and epoxymycolates and both serovars of Mycobacterium simiae had a very characteristic pattern of -, - and ketomycolic acids. Comparison with data for other mycobacteria showed the chemotaxonomic significance of these mycolic acid patterns.Abbreviations TLC
thin-layer chromatography
- TBDMS
t-butyldimethylsilyl
Supplementary data: Copies of the chromatographic patterns of the mycolic acids from all the strains examined can be provided on request from one of the authors (D.E.M.) 相似文献
12.
Philippe Pelupessy Elisabetta Chiarparin Ranajeet Ghose Geoffrey Bodenhausen 《Journal of biomolecular NMR》1999,13(4):375-380
The angle CH,NHN subtended by the internuclear vectors 13C-H and 15N-HN in doubly-labeled proteins can be determined by observing the effect of cross-correlation between the dipolar interactions on zero- and double-quantum coherences involving 13C and 15N. Two complementary 2D experiments with the appearance of 15N-HN correlation spectra yield signal intensities that depend on the rate of interconversion through cross-correlated relaxation of in-phase and doubly antiphase zero- and double-quantum coherences. The ratio of the signal intensities in the two experiments bears a simple relationship to the cross-correlation rate, and hence to the angle CH,NHN. Assuming planarity of the peptide bond, the dihedral angle (between C and C) can be determined from the knowledge of CH,NHN. The experiments are very time-effective and provide good sensitivity and excellent spectral resolution. 相似文献
13.
Eric L. Charnov 《Evolutionary ecology》1989,3(3):236-239
Summary Darwinian fitness with respect to the age of maturity () in stationary populations can be written as the product of two functions: pre- survival times the Fisherian reproductive value of an age (a just mature) individual. This reduces normalizing selection on to the maximization of a simple product,a twodimensional problem (Charnov in press). I apply this products theorem to in Pandalid shrimp and compare the results to previous analysis (Roff, 1986) of in fish. 相似文献
14.
Ornithine decarboxylase prevents tumor necrosis factor alpha-induced apoptosis by decreasing intracellular reactive oxygen species 总被引:1,自引:0,他引:1
Liu GY Hung YC Hsu PC Liao YF Chang WH Tsay GJ Hung HC 《Apoptosis : an international journal on programmed cell death》2005,10(3):569-581
Ornithine decarboxylase (ODC) plays an essential role in various biological functions, including cell proliferation, differentiation and cell death. However, how it prevents the cell apoptotic mechanism is still unclear. Previous studies have demonstrated that decreasing the activity of ODC by difluoromethylornithine (DFMO), an irreversible inhibitor of ODC, causes the accumulation of intracellular reactive oxygen species (ROS) and cell arrest, thus inducing cell death. These findings might indicate how ODC exerts anti-oxidative and anti-apoptotic effects. In our study, tumor necrosis factor alpha (TNF-) induced apoptosis in HL-60 and Jurkat T cells. The kinetic studies revealed that the TNF- -induced apoptotic process included intracellular ROS generation (as early as 1 h after treatment), the activation of caspase 8 (3 h), the cleavage of Bid (3 h) and the disruption of mitochondrial membrane potential ( m) (6 h). Furthermore, ROS scavengers, such as glutathione (GSH) and catalase, maintained m and prevented apoptosis upon treatment. Putrescine and overexpression of ODC had similar effects as ROS scavengers in decreasing intracellular ROS and preventing the disruption of m and apoptosis. Inhibition of ODC by DFMO in HL-60 cells only could increase ROS generation, but did not disrupt m or induce apoptosis. However, DFMO enhanced the accumulation of ROS, disruption of m and apoptosis when cells were treated with TNF- . ODC overexpression avoided the decline of Bcl-2, prevented cytochrome c release from mitochondria and inhibited the activation of caspase 8, 9 and 3. Overexpression of Bcl-2 maintained m and prevented apoptosis, but could not reduce ROS until four hours after TNF- treatment. According to these data, we suggest that TNF- induces apoptosis mainly by a ROS-dependent, mitochondria-mediated pathway. Furthermore, ODC prevents TNF- -induced apoptosis by decreasing intracellular ROS to avoid Bcl-2 decline, maintain m, prevent cytochrome c release and deactivate the caspase cascade pathway. 相似文献
15.
Donor tissue and culture condition effects on mesophyll protoplasts of Medicago sativa 总被引:6,自引:0,他引:6
Mesophyll protoplasts were produced from clones of two cultivars of Medicago sativa, Rangelander and Regen S. Protoplasts from the Regen S clone generally gave rise to calli while those from the Rangelander clone would undergo direct embryogenesis. Effects of plant growth conditions, donor tissue pretreatment and protoplast culture conditions on mesophyll protoplast production and subsequent development patterns were investigated. The major factor determining whether or not mesophyll protoplasts would be produced from either of the clones was the pretreatment in water of shoots excised from the donor plants. Pretreatment in water containing growth regulators did not alter protoplast production or development in the Regen S clone. Pretreatment of the Rangelander clone shoots with abscisic acid or naphthaleneacetic acid was slightly beneficial to embryo production while pretreatment with benzylaminopurine was detrimental. Altered leaf morphology induced by growth condition changes did not affect mesophyll protoplast production or subsequent development patterns when shoots were pretreated in water. Culture of protoplasts in liquid droplets or solid agar medium increased low density protoplast survival and subsequent embryo production in the Rangelander clone. 相似文献
16.
The genes encoding three invariant components of the human T-cell antigen receptor, the CD3 , , and chains, are located on human chromosome 11 at band q23. We isolated cosmid clones containing the human CD3 and chain genes in vectors designed for rapid and efficient chromosome walking. The human CD3 gene was located in the region immediately downstream of the CD3 and genes using synthetic oligonucleotide probes and the localization of this gene confirmed by DNA sequencing. Detailed restriction mapping of the CD3 locus demonstrated that all three CD3 subunits are encoded within 60 kb of DNA with the CD3 gene located 26 kb downstream of the CD3 and genes. Analysis of genomic DNA on pulsed field gels using probes isolated from these cosmid clones defined a physical map of 750 kb spanning the CD3 locus on human chromosome 11g23. The CD3 genes thus comprise a multigene family encoding cell surface components important for transmembrane signaling on T lymphocytes. The arrangement of these genes suggest that they may share common regulatory elements for the control of gene expression during T-cell ontogeny. 相似文献
17.
The growth of miniature rose (Rosa chinensis Jacq. Minima) shoots cultured on liquid medium was greater relative to those cultured on two-phase (solid + liquid) medium or solid medium alone. Shoot multiplication ratio (number of multiple shoots per explant per subculture) on liquid medium was higher with 17.8–26.6 M 6-benzyladenine at compared to that at 0–8.9 M. Shoots grown on 30 ml or more of liquid medium had a higher multiplication ratio than those grown on 10 or 20 ml. The growth and multiplication ratio increased when the culture period was extended from 3 to 6 weeks, although plantlets began to exhibit some chlorosis by the 6th week. These conditions were maintained over four subcultures for cultivars Baby Katie, Lavender Jewel, Red Sunblaze and Royal Sunblaze, with no significant change in multiplication ratio over time.Abbreviations BA
6-benzyladenine
- NAA
1-naphthaleneacetic acid 相似文献
18.
Mark A. Williams Hiroshi Kitagawa Arun K. Datta James C. Paulson James C. Jamieson 《Glycoconjugate journal》1995,12(6):755-761
Values ofK
m were determined for three purified sialyltransferases and the corresponding recombinant enzymes. The enzymes were Gal1-4GlcNAc 2-6sialyltransferase and Gal1-3(4)GlcNAc 2-3sialyltransferase from rat liver; these enzymes are responsible for the attachment of sialic acid to N-linked oligosaccharide chains; and the Gal1-3GalNAc 2-3sialyltransferase from porcine submaxillary gland that is responsible for the attachment of sialic acid to O-linked glycoproteins and glycolipids. A procedure for the large scale expression of active sialyltransferases from recombinant baculovirus-infected insect cells is described. For the liver enzymes values ofK
m were determined using rat and human asialo1 acid glycoprotein andN-acetyllactosamine as variable substrates; lacto-N-tetraose was also used with the Gal1-3(4)GlcNAc 2-3sialyltransferase. Antifreeze glycorprotein was used as the macromolecular acceptor for the porcine enzyme. Values forK
m were also determined using CMP-NeuAc as the variable substrate.Abbreviations NeuAc
N-acetylneuraminic acid
- Gal
galactose
- GlcNAc
N-acetylglucosamine
Enzymes: Gal1-4GlcNAc 2-6sialyltransferase, EC 2.4.99.1; Gal1-3(4)GlcNAc 2-3sialyltransferase, EC 2.4.99.5; Gal1-3GalNAc 2-3sialyltransferase, EC 2.4.99.4. 相似文献
19.
This paper gives, for the first time, a complete history of rain forest canopy access, from the early years to the present day. This review is primarily from the European perspective, and explores the development of canopy access techniques, from low-tech methods such as single rope technique, to hi-tech approaches such as canopy cranes. In recent years, canopy science has moved away from pure exploration (the Wonderland phase) to tackling the practicalities of rigorous canopy research (Reality), and the underlying emphasis is now shifting from access to the upper canopy per se to conducting replicative and manipulative science. The paper concludes by advocating the integration of many access techniques (both hi-tech and low-tech) at selected research sites, and certain neglected key areas of research are highlighted, including the comparison of adjacent primary and logged forests. 相似文献
20.
Jeffrey M. Leiden Keith M. Gottesdiener Thomas Quertermous Larry Coury Robert A. Bray Lisa Gottschalk Howard Gebel Jon G. Seidman Jack L. Strominger Alan L. Landay Jacki Kornbluth 《Immunogenetics》1988,27(4):231-238
To test the hypothesis that the T-cell receptor (Tcr) gene encodes a natural killer (NK) cell receptor molecule, three human NK clones and fresh peripheral blood lymphocytes with NK activity from two patients with a CD16+ lymphocytosis were analyzed for rearrangements and expression of the human Tcr , , and genes. Two of the clones displayed distinct rearrangements of their Tcr and genes and expressed mature Tcr , , and l RNA. However, one of the clones and both patient samples displayed marked NK activity but failed to rearrange or express any of their Tcr genes. These findings demonstrate that human natural killer activity is not dependent on Tcr gene rearrangement and expression. In addition, they confirm previous findings concerning the lack of Tcr and gene expression in some natural killer cells. Thus, they suggest the existence of additional NK-specific recognition molecules. 相似文献