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1.
Occurrence of Bacillus subtilis with High Heat Resistance 总被引:4,自引:3,他引:1
2.
An De Bondt Kristel Eggermont Iris Penninckx Inge Goderis Willem F. Broekaert 《Plant cell reports》1996,15(7):549-554
We have previously developed a protocol for efficient gene transfer and regeneration of transgenic calli following cocultivation of apple (cv. Jonagold) explants with Agrobacterium tumefaciens (De Bondt et al. 1994, Plant Cell Reports 13: 587–593). Now we report on the optimization of postcultivation conditions for efficient and reproducible regeneration of transgenic shoots from the apple cultivar Jonagold. Factors which were found to be essential for efficient shoot regeneration were the use of gelrite as a gelling agent and the use of the cytokinin-mimicing thidiazuron in the selective postcultivation medium. Improved transformation efficiencies were obtained by combining the hormones thidiazuron and zeatin and by using leaf explants from in vitro grown shoots not older than 4 weeks after multiplication. Attempts to use phosphinothricin acetyl transferase as a selectable marker were not successful. Using selection on kanamycin under optimal postcultivation conditions, about 2% of the leaf explants developed transgenic shoots or shoot clusters. The presence and expression of the transferred genes was verified by -glucuronidase assays and Southern analysis. The transformation procedure has also been succesfully applied to several other apple cultivars.Abbreviations BAP
benzylaminopurine
- CTAB
hexadecyltrimethylammoniumbromide
- Na2EDTA
ethylenediamine-tetra-acetate ferric-sodium salt
- FeNaEDTA
ethylenediamine-tetra-acetate ferric-sodium salt
- GA3
gibberellic acid 3
- GusA
-glucuronidase
-
gusA
-glucuronidase gene of Escherichia coli
- IAA
indole acetic acid
- IBA
indole butyric acid
- 2iP
N6-2-isopentenyl adenine
- NAA
naphthalene acetic acid
-
nptII
neomycinphosphotransferase II gene
-
bar
phosphinothricin acetyl transferase gene
- PCR
polymerase chain reaction
- PPT
phosphinothricin
- STS
silver thiosulphate
- T-DNA
transferred DNA
- TDZ
thidiazuron
- X-Gluc
5-bromo-4-chloro-3-indolyl -D-glucuronide
- Zea
trans-Zeatin 相似文献
3.
Kristel Eggermont Inge J. Goderis Willem F. Broekaert 《Plant Molecular Biology Reporter》1996,14(3):273-279
We describe here a reliable high-throughput method for extraction of RNA from fresh or frozen plant tissue that obviates laborious
and time-consuming homogenisation by mortar and pestle. The method is based on homogenisation by high-speed reciprocal shaking
in presence of a mixture of inexpensive abrasive materials; i.e., quartz sand and glass beads. After homogenisation, the method
follows a standard procedure for RNA extraction by phenol/LiCl. Yield and quality of RNA obtained by homogenisation with the
sand/glass bead mix are identical to those obtained by mortar and pestle. 相似文献
4.
Mobility during the neolithic and bronze age in northern ireland explored using strontium isotope analysis of cremated human bone
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5.
For lintners with negligible amylose retrogradation, crystallinity related inversely to starch amylose content and, irrespective of starch source, incomplete removal of amorphous material was shown. The latter was more pronounced for B-type than for A-type starches. The two predominant lintner populations, with modal degrees of polymerization (DP) of 13-15 and 23-27, were best resolved for amylose-deficient and A-type starches. Results indicate a more specific hydrolysis of amorphous lamellae in such starches. Small-angle X-ray scattering showed a more intense 9-nm scattering peak for native amylose-deficient A-type starches than for their regular or B-type analogues. The experimental evidence indicates a lower contrasting density within the "crystalline" shells of the latter starches. A higher density in the amorphous lamellae, envisaged by the lamellar helical model, explains the relative acid resistance of linear amylopectin chains with DP > 20, observed in lintners of B-type starches. Because amylopectin chain length distributions were similar for regular and amylose-deficient starches of the same crystal type, we deduce that the more dense (and ordered) packing of double helices into lamellar structures in amylose-deficient starches is due to a different amylopectin branching pattern. 相似文献
6.
L. Verdoodt A. Van Haute I. J. Goderis K. De Witte J. Keulemans W. Broothaerts 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1998,96(2):294-300
To obtain homozygous genotypes of apple, we have induced haploid development of either the female or the male gametes by
parthenogenesis in situ and anther culture, respectively. Of the shoots obtained, which were mainly of a non-haploid nature,
some could be derived from fertilised egg cells or from sporophytic anther tissue. In order to select the shoots having a
true haploid origin, and thus homozygotes, we decided to use the single multi-allelic self-incompatibility gene as a molecular
marker to discriminate homozygous from heterozygous individuals. The rationale behind this approach was that diploid apple
cultivars contain 2 different alleles of the S-gene and therefore the haploid induced shoots obtained from them should have only one of the alleles of the single parent.
The parental cultivars used were ‘Idared’ (parthenogenesis in situ) and ‘Braeburn’ (androgenesis), and their S-genotypes were known, except for 1 of the ‘Braeburn’S-alleles. To stimulate parthenogenetic development ‘Idared’ styles were pollinated with irradiated ‘Baskatong’ pollen, the
S-alleles of the latter (2n) cultivar were also unknown. The cloning and sequence analysis of these 3 unidentified S-alleles, 1 from ‘Braeburn’ and 2 from ‘Baskatong’ is described, and we show that they correspond to the S
24
-, S
26
- and S
27
-alleles. We have optimised a method for analysis of the S-alleles of ‘Idared/Baskatong’- or ‘Braeburn’-derived in vitro plant tissues and have shown that this approach can be applied
for the screening of the in vitro shoots for their haploid origin.
Received: 18 August 1997 / Accepted: 10 September 1997 相似文献
7.
Purification and characterization of Dolichos lablab lectin 总被引:1,自引:0,他引:1
The mannose/glucose-binding Dolichos lablab lectin (designated DLL) has
been purified from seeds of Dolichos lablab (hyacinth bean) to
electrophoretic homogeneity by affinity chromatography on an ovalbumin-
Sepharose 4B column. The purified lectin gave a single symmetric protein
peak with an apparent molecular mass of 67 kDa on gel filtration
chromatography, and five bands ranging from 10 kDa to 22 kDa upon SDS-PAGE.
N-Terminal sequence analysis of these bands revealed subunit heterogeneity
due to posttranslational proteolytic truncation at different sites mostly
at the carboxyl terminus. The carbohydrate binding properties of the
purified lectin were investigated by three different approaches:
hemagglutination inhibition assay, quantitative precipitation inhibition
assay, and ELISA. On the basis of these studies, it is concluded that the
Dolichos lablab lectin has neither an extended carbohydrate combining site,
nor a hydrophobic binding site adjacent to it. The carbohydrate combining
site of DLL appears to most effectively accommodate a nonreducing terminal
alpha-d-mannosyl unit, and to be complementary to the C-3, C-4, and C-6
equatorial hydroxyl groups of alpha-d-mannopyranosyl and
alpha-d-glucopyranosyl residues. DLL strongly precipitates murine IgM but
not IgG, and the recent finding that this lectin interacts specifically
with NIH 3T3 fibroblasts transfected with the Flt3 tyrosine kinase receptor
and preserves human cord blood stem cells and progenitors in a quiescent
state for prolonged periods in culture, make this lectin a valuable tool in
biomedical research.
相似文献
8.
不同胁迫预处理提高水稻幼苗抗寒性期间蛋白质的变化 总被引:13,自引:0,他引:13
水稻(Oryza sativa L.)幼苗经盐、热激和冷三种不同胁迫预处理均提高了幼苗的抗寒性。与未预处理苗相比,在处理后、低温伤害后和常温下恢复2d的三个时期,不同胁迫预处理苗的可溶性和热不稳定蛋白含量变化趋势甚为相似,但热稳定蛋白含量变化则各有异同。SDS-PAGE图谱分析显示,不同胁迫预处理提高水稻幼苗抗寒性时,其可溶性蛋白、热稳定和热不稳定蛋白组成变化亦各有异同。除诱导出共有的新多肽外,还各自诱导出特有的新多肽。结果表明,植物对不同胁迫的交叉适应存在一定的共同机理,但亦可看出植物对同一种环境胁迫似乎不是以同一的机理去适应。 相似文献
9.
Molecular and morphological aspects of annealing-induced stabilization of starch crystallites 总被引:5,自引:0,他引:5
Gomand SV Lamberts L Gommes CJ Visser RG Delcour JA Goderis B 《Biomacromolecules》2012,13(5):1361-1370
A unique series of potato (mutant) starches with highly different amylopectin/amylose (AP/AM) ratios was annealed in excess water at stepwise increasing temperatures to increase the starch melting (or gelatinization) temperatures in aqueous suspensions. Small-angle X-ray scattering (SAXS) experiments revealed that the lamellar starch crystals gain stability upon annealing via thickening for high-AM starch, whereas the crystal surface energy decreases for AM-free starch. In starches with intermediate AP/AM ratio, both mechanisms occur, but the surface energy reduction mechanism prevails. Crystal thickening seems to be associated with the cocrystallization of AM with AP, leading to very disordered nanomorphologies for which a new SAXS data interpretation scheme needed to be developed. Annealing affects neither the crystal internal structure nor the spherulitic morphology on a micrometer length scale. 相似文献
10.
Small unilamellar vesicles were used to measure the permeability of saturated phosphatidylcholine bilayers to glucose. The presented method circumvents most of the common restrictions of classical permeability experiments. Increasing the fatty acid chain length of the lipids reduced the permeation rate significantly. Raising the temperature above that of the lipid phase transition drastically increased membrane permeability. Arrhenius plots demonstrated the activation energy to be independent of membrane composition and the phase-state of the lipids. The permeation process is discussed in terms of a constant energy to disrupt all hydrogen bonds between permeant and aqueous solvent prior to penetrating the membrane. The magnitude of the permeability coefficient is partly determined by a unfavourable change in entropy of activation on crossing the water/lipid interface. All results indicate that the penetration of the dehydrated permeant into the hydrophobic barrier is the rate-limiting step in the permeation of glucose. 相似文献