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161.
Martin M. Kater Gregory M. Koningstein H. John J. Nijkamp Antoine R. Stuitje 《Plant molecular biology》1994,25(5):771-790
Fatty acid synthesis in bacteria and plants is catalysed by a multi-enzyme fatty acid synthetase complex (FAS II) which consists of separate monofunctional polypeptides. Here we present a comparative molecular genetic and biochemical study of the enoyl-ACP reductase FAS components of plant and bacterial origin. The putative bacterial enoyl-ACP reductase gene (envM) was identified on the basis of amino acid sequence similarities with the recently cloned plant enoyl-ACP reductase. Subsequently, it was unambiguously demonstrated by overexpression studies that theenvM gene encodes the bacterial enoyl-ACP reductase. An anti-bacterial agent called diazaborine was shown to be a specific inhibitor of the bacterial enoyl-ACP reductase, whereas the plant enzyme was insensitive to this synthetic antibiotic. The close functional relationship between the plant and bacterial enoyl-ACP reductases was inferred from genetic complementation of anenvM mutant ofEscherichia coli. Ultimately,envM gene-replacement studies, facilitated by the use of diazaborine, demonstrated for the first time that a single component of the plant FAS system can functionally replace its counterpart within the bacterial multienzyme complex. Finally, lipid analysis of recombinantE. coli strains with the hybrid FAS system unexpectedly revealed that enoyl-ACP reductase catalyses a rate-limiting step in the elongation of unsaturated fatty acids. 相似文献
162.
Se Ryun Kwon Yue Jai Kang Dong Jin Lee Eun Hye Lee Yoon Kwon Nam Sung Koo Kim Ki Hong Kim 《Molecular biotechnology》2009,42(2):154-159
Vibrio anguillarum ghosts (VAG) were generated, for the first time, using a conjugation vector containing a ghost bacteria inducing cassette,
pRK-λPR-cI-Elysis, in which the expression of PhiX174 lysis gene E was controlled by the P
R
/cI regulatory system of lambda phage. By scanning electron microscopy, holes ranging 80–200 nm in diameter were observed in
the VAG. To avoid the presence of bacterial genomic DNA and an antibiotic resistance gene in the final VAG product, we constructed
a new dual vector, pRK-λPR-cI-E-SNA, containing the E-mediated lysis cassette and the staphylococcal nuclease A (SNA)-mediated DNA degradation cassette, and generated safety-enhanced VAG for use as a fish vaccine. 相似文献
163.
Kai-Shu Ling Karen R. Harris Jenelle D. F. Meyer Amnon Levi Nihat Guner Todd C. Wehner Abdelhafid Bendahmane Michael J. Havey 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2009,120(1):191-200
Zucchini yellow mosaic virus (ZYMV) is one of the most economically important potyviruses infecting cucurbit crops worldwide. Using a candidate gene approach,
we cloned and sequenced eIF4E and eIF(iso)4E gene segments in watermelon. Analysis of the nucleotide sequences between the
ZYMV-resistant watermelon plant introduction PI 595203 (Citrullus lanatus var. lanatus) and the ZYMV-susceptible watermelon cultivar ‘New Hampshire Midget’ (‘NHM’) showed the presence of single nucleotide polymorphisms
(SNPs). Initial analysis of the identified SNPs in association studies indicated that SNPs in the eIF4E, but not eIF(iso)4E,
were closely associated to the phenotype of ZYMV-resistance in 70 F2 and 114 BC1R progenies. Subsequently, we focused our efforts in obtaining the entire genomic sequence of watermelon eIF4E. Three SNPs
were identified between PI 595203 and NHM. One of the SNPs (A241C) was in exon 1 and the other two SNPs (C309A and T554G)
were in the first intron of the gene. SNP241 which resulted in an amino acid substitution (proline to threonine) was shown
to be located in the critical cap recognition and binding area, similar to that of several plant species resistance to potyviruses.
Analysis of a cleaved amplified polymorphism sequence (CAPS) marker derived from this SNP in F2 and BC1R populations demonstrated a cosegregation between the CAPS-2 marker and their ZYMV resistance or susceptibility phenotype.
When we investigated whether such SNP mutation in the eIF4E was also conserved in several other PIs of C. lanatus var. citroides, we identified a different SNP (A171G) resulting in another amino acid substitution (D71G) from four ZYMV-resistant C. lanatus var. citroides (PI 244018, PI 482261, PI 482299, and PI 482322). Additional CAPS markers were also identified. Availability of all these
CAPS markers will enable marker-aided breeding of watermelon for ZYMV resistance. 相似文献
164.
Ascorbate (AsA) plays a fundamental role in redox homeostasis in plants and animals, primarily by scavenging reactive oxygen
species. Three genes, representing diverse steps putatively involved in plant AsA biosynthesis pathways, were cloned and independently
expressed in Solanum lycopersicum (tomato) under the control of the CaMV 35S promoter. Yeast-derived GDP-mannose pyrophosphorylase (GMPase) and arabinono-1,4-lactone oxidase (ALO), as well as myo-inositol oxygenase 2 (MIOX2) from Arabidopsis thaliana, were targeted. Increases in GMPase activity were concomitant with increased AsA levels of up to 70% in leaves, 50% in green
fruit, and 35% in red fruit. Expression of ALO significantly pulled biosynthetic flux towards AsA in leaves and green fruit by up to 54 and 25%, respectively. Changes in
AsA content in plants transcribing the MIOX2 gene were inconsistent in different tissue. On the other hand, MIOX activity was strongly correlated with cell wall uronic
acid levels, suggesting that MIOX may be a useful tool for the manipulation of cell wall composition. In conclusion, the Smirnoff–Wheeler
pathway showed great promise as a target for biotechnological manipulation of ascorbate levels in tomato. 相似文献
165.
Four different bacterial isolates obtained from a stable bacterial consortium were capable of utilizing pentachlorophenol
(PCP) as sole carbon and energy source. The consortium was developed by continuous enrichment in the chemostat. The degradation
of PCP by bacterial strain was preceded through an oxidative route as indicated by accumulation of tetrachloro-ρ-hydroquinone
and dichlorohydroquinone as determined by high performance liquid chromatography (HPLC). Among the four isolates, Pseudomonas fluorescens exhibited maximum degradation capability and enzyme production. PCP-monooxygenase enzyme was extracted from culture extract
and fractionated by DEAE-cellulose ion exchange chromatography. The molecular weight of the enzyme, purified from Pseudomonas fluorescens, determined by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and gel filtration chromatography was
found to be 24,000 Da.
Received: 22 July 2002 / Accepted: 23 September 2002 相似文献
166.
Hernández-Hernández A Rincón-Arano H Recillas-Targa F Ortiz R Valdes-Quezada C Echeverría OM Benavente R Vázquez-Nin GH 《Chromosoma》2008,117(1):77-87
The synaptonemal complex (SC) is an evolutionarily conserved structure that mediates synapsis of homologous chromosomes during
meiotic prophase I. Previous studies have established that the chromatin of homologous chromosomes is organized in loops that
are attached to the lateral elements (LEs) of the SC. The characterization of the genomic sequences associated with LEs of
the SC represents an important step toward understanding meiotic chromosome organization and function. To isolate these genomic
sequences, we performed chromatin immunoprecipitation assays in rat spermatocytes using an antibody against SYCP3, a major
structural component of the LEs of the SC. Our results demonstrated the reproducible and exclusive isolation of repeat deoxyribonucleic
acid (DNA) sequences, in particular long interspersed elements, short interspersed elements, long terminal direct repeats,
satellite, and simple repeats. The association of these repeat sequences to the LEs of the SC was confirmed by in situ hybridization
of meiotic nuclei shown by both light and electron microscopy. Signals were also detected over the chromatin surrounding SCs
and in small loops protruding from the lateral elements into the SC central region. We propose that genomic repeat DNA sequences
play a key role in anchoring the chromosome to the protein scaffold of the SC.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. 相似文献
167.
The promoter of the pepper pathogen-induced membrane protein gene CaPIMP1 was analyzed by an Agrobacterium-mediated transient expression assay in tobacco leaves. Several stress-related cis-acting elements (GT-1, W-box and ABRE) are located within the CaPIMP1 promoter. In tobacco leaf tissues transiently transformed with a CaPIMP1 promoter-β-glucuronidase (GUS) gene fusion, serially 5′-deleted CaPIMP1 promoters were differentially activated by Pseudomonas syringae pv. tabaci, ethylene, methyl jasmonate, abscisic acid, and nitric oxide. The −1,193 bp region of the CaPIMP1 gene promoter sequence exhibited full promoter activity. The −417- and −593 bp promoter regions were sufficient for GUS gene activation by ethylene and methyl jasmonate treatments, respectively. However, CaPIMP1 promoter sequences longer than −793 bp were required for promoter activation by abscisic acid and sodium nitroprusside treatments.
CaPIMP1 expression was activated in pepper leaves by treatment with ethylene, methyl jasmonate, abscisic acid, β-amino-n-butyric acid, NaCl, mechanical wounding, and low temperature, but not with salicylic acid. Overexpression of CaPIMP1 in Arabidopsis conferred hypersensitivity to mannitol, NaCl, and ABA during seed germination but not during seedling development. In contrast,
transgenic plants overexpressing CaPIMP1 exhibited enhanced tolerance to oxidative stress induced by methyl viologen during germination and early seedling stages.
These results suggest that CaPIMP1 expression may alter responsiveness to environmental stress, as well as to pathogen infection.
The nucleotide sequence data reported here has been deposited in the GenBank database under the accession number DQ356279. 相似文献
168.
Nitric oxide (NO) is an important molecule that acts in many tissues to regulate a diverse range of physiological processes.
It is becoming apparent that NO is a ubiquitous signal in plants. Since the discovery of NO emission by plants in the 1970s,
this gaseous compound has emerged as a major signalling molecule involved in multiple physiological functions. Research on
NO in plants has gained significant awareness in recent years and there is increasing indication on the role of this molecule
as a key-signalling molecule in plants. The investigations about NO in plants have been concentrated on three main fields:
The search of NO or any source of NO generation, effects of exogenous NO treatments, NO transduction pathways. However we
have limited information about signal transduction procedures by which NO interaction with cells results in altered cellular
activities. This article reviews recent advances in NO synthesis and its signalling functions in plants. First, different
sources and biosynthesis of NO in plants, then biological processes involving NO signalling are reviewed. NO signalling relation
with cGMP, protein kinases and programmed cell death are also discussed. Besides, NO signalling in plant defense response
is also examined. Especially NO signalling between animal and plant systems is compared. 相似文献
169.
Hisashi Mizutani Hideaki Sugawara Ashley M. Buckle Takeshi Sangawa Ken-ichi Miyazono Jun Ohtsuka Koji Nagata Tomoki Shojima Shohei Nosaki Yuqun Xu Delong Wang Xiao Hu Masaru Tanokura Kei Yura 《BMC structural biology》2017,17(1):4
Background
More than 7000 papers related to “protein refolding” have been published to date, with approximately 300 reports each year during the last decade. Whilst some of these papers provide experimental protocols for protein refolding, a survey in the structural life science communities showed a necessity for a comprehensive database for refolding techniques. We therefore have developed a new resource – “REFOLDdb” that collects refolding techniques into a single, searchable repository to help researchers develop refolding protocols for proteins of interest.Results
We based our resource on the existing REFOLD database, which has not been updated since 2009. We redesigned the data format to be more concise, allowing consistent representations among data entries compared with the original REFOLD database. The remodeled data architecture enhances the search efficiency and improves the sustainability of the database. After an exhaustive literature search we added experimental refolding protocols from reports published 2009 to early 2017. In addition to this new data, we fully converted and integrated existing REFOLD data into our new resource. REFOLDdb contains 1877 entries as of March 17th, 2017, and is freely available at http://p4d-info.nig.ac.jp/refolddb/.Conclusion
REFOLDdb is a unique database for the life sciences research community, providing annotated information for designing new refolding protocols and customizing existing methodologies. We envisage that this resource will find wide utility across broad disciplines that rely on the production of pure, active, recombinant proteins. Furthermore, the database also provides a useful overview of the recent trends and statistics in refolding technology development.170.
The Effect of Sterilization Methods on the Physical Properties of Silk Sericin Scaffolds 总被引:1,自引:0,他引:1
Protein-based biomaterials respond differently to sterilization methods. Since protein is a complex structure, heat, or irradiation
may result in the loss of its physical or biological properties. Recent investigations have shown that sericin, a degumming
silk protein, can be successfully formed into a 3-D scaffolds after mixing with other polymers which can be applied in skin
tissue engineering. The objective of this study was to investigate the effectiveness of ethanol, ethylene oxide (EtO) and
gamma irradiation on the sterilization of sericin scaffolds. The influence of these sterilization methods on the physical
properties such as pore size, scaffold dimensions, swelling and mechanical properties, as well as the amount of sericin released
from sericin/polyvinyl alcohol/glycerin scaffolds, were also investigated. Ethanol treatment was ineffective for sericin scaffold
sterilization whereas gamma irradiation was the most effective technique for scaffold sterilization. Moreover, ethanol also
caused significant changes in pore size resulting from shrinkage of the scaffold. Gamma-irradiated samples exhibited the highest
swelling property, but they also lost the greatest amount of weight after immersion for 24 h compared with scaffolds obtained
from other sterilization methods. The results of the maximum stress test and Young’s modulus showed that gamma-irradiated
and ethanol-treated scaffolds are more flexible than the EtO-treated and untreated scaffolds. The amount of sericin released,
which was related to its collagen promoting effect, was highest from the gamma-irradiated scaffold. The results of this study
indicate that gamma irradiation should have the greatest potential for sterilizing sericin scaffolds for skin tissue engineering. 相似文献