Abstract:
:Here, we provide comprehensive, highly efficient protocols for Agrobacterium tumefaciens-mediated transformation of a wide range of rice genotypes. Methods that use either immature embryos (japonica and indica rice) or calli (japonica cultivars and the indica cultivar, Kasalath) as a starting material for inoculation with Agrobacterium are described. Immature embryos are pretreated with heat and centrifugal force, which significantly enhances the efficiency of gene transfer, and then infected with Agrobacterium. Callus is induced from mature seeds and infected. Transformed cells proliferated from these tissues are selected on the basis of hygromycin resistance, and transgenic plants are eventually regenerated. A single immature japonica or Kasalath embryo will produce between 10 and 18 independent transgenic plants; for other non-Kasalath indica varieties, the number of transgenic plants expected will be between 5 and 13. For japonica and Kasalath, transformants should be obtained from between 50 and 90% of calli. From inoculation with Agrobacterium to transplanting to soil will take 55 d for japonica and Kasalath, and 74 d for indica other than Kasalath using the immature embryo method, and 50 d for japonica and Kasalath using the callus method.
journal_name
Nat Protocjournal_title
Nature protocolsauthors
Hiei Y,Komari Tdoi
10.1038/nprot.2008.46subject
Has Abstractpub_date
2008-01-01 00:00:00pages
824-34issue
5eissn
1754-2189issn
1750-2799pii
nprot.2008.46journal_volume
3pub_type
杂志文章相关文献
Nature Protocols文献大全abstract::'Adult' or 'somatic' stem cells harbor an intrinsic ability to regenerate tissues. Heterogeneity of such stem cells along the gastrointestinal tract yields the known segmental specificity of this organ and may contribute to the pathology of certain enteric conditions. Here we detail technology for the generation of 'l...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/s41596-020-0298-4
更新日期:2020-05-01 00:00:00
abstract::We present a protocol for visualizing and quantifying single mRNA molecules in mammalian (mouse and human) tissues. In the approach described here, sets of about 50 short oligonucleotides, each labeled with a single fluorophore, are hybridized to target mRNAs in tissue sections. Each set binds to a single mRNA molecul...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2013.109
更新日期:2013-09-01 00:00:00
abstract::Here, we describe a simple protocol for the design and construction of a laser-guided direct writing (LGDW) system able to micropattern the self-assembly of liver sinusoid-like structures with micrometer resolution in vitro. To the best of our knowledge, LGDW is the only technique able to pattern cells "on the fly" wi...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2006.386
更新日期:2006-01-01 00:00:00
abstract::The dynamics of transcription can be studied genome wide by high-throughput sequencing of nascent and newly synthesized RNA. 4-thiouridine (4SU) labeling in vivo enables the specific capture of such new transcripts, with 4SU residues being tagged by biotin linkers and captured using streptavidin beads before library p...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/s41596-019-0262-3
更新日期:2020-02-01 00:00:00
abstract::The cytokinesis-block micronucleus cytome assay is a comprehensive system for measuring DNA damage, cytostasis and cytotoxicity. DNA damage events are scored specifically in once-divided binucleated (BN) cells and include (a) micronuclei (MNi), a biomarker of chromosome breakage and/or whole chromosome loss, (b) nucle...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2007.77
更新日期:2007-01-01 00:00:00
abstract::This protocol describes an in vitro approach for measuring the kinetics and affinities of interactions between membrane-anchored proteins. This method is particularly established for dissecting the interaction dynamics of cytokines with their receptor subunits. For this purpose, the receptor subunits are tethered in a...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2006.270
更新日期:2006-01-01 00:00:00
abstract::Neurotransmitter release is triggered by membrane depolarization, Ca(2+) influx and Ca(2+) sensing by the release machinery, causing synaptic vesicle (SV) fusion with the plasma membrane. Interlinked is a complex membrane cycle in which vesicles are tethered to the release site, primed, fused and recycled. As many of ...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2012.074
更新日期:2012-06-21 00:00:00
abstract::Cyclodepsipeptides are cyclic peptides in which at least one amide link on the backbone is replaced with an ester link. These natural products present a high structural diversity that corresponds to a broad range of biological activities. Therefore, they are very promising pharmaceutical candidates. Most of the cyclod...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2016.116
更新日期:2016-10-01 00:00:00
abstract::The immune system operates at the scale of the whole organism in mammals. We currently lack experimental approaches to systematically track and study organism-wide molecular processes in mice. Here we describe an integrated toolkit for measuring gene expression in whole tissues, 3-prime mRNA extension sequencing, that...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/s41596-019-0291-y
更新日期:2020-04-01 00:00:00
abstract::Efforts to cure HIV are hampered by limited characterization of the cells supporting HIV replication in vivo and inadequate methods for quantifying the latent viral reservoir in individuals receiving antiretroviral therapy (ART). We describe a protocol for flow cytometric identification of viral reservoirs, based on c...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2017.079
更新日期:2017-10-01 00:00:00
abstract::Boronic acids and esters have critical roles in the areas of synthetic organic chemistry, molecular sensors, materials science, drug discovery, and catalysis. Many of the current applications of boronic acids and esters require materials with very low levels of transition metal contamination. Most of the current metho...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2016.184
更新日期:2017-03-01 00:00:00
abstract::Small-angle X-ray scattering (SAXS) and small-angle neutron scattering (SANS) are techniques used to extract structural parameters and determine the overall structures and shapes of biological macromolecules, complexes and assemblies in solution. The scattering intensities measured from a sample contain contributions ...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2016.113
更新日期:2016-11-01 00:00:00
abstract::This protocol describes a simple means of measuring the starch content of plant tissues by solubilizing the starch, converting it quantitatively to glucose and assaying the glucose. Plant tissue must initially be frozen rapidly to stop metabolism, then extracted to remove free glucose. Starch is solubilized by heating...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2006.232
更新日期:2006-01-01 00:00:00
abstract::This protocol describes an optimized method for direct in vitro monitoring of homo- and heterotypic axon-axon interactions involved in the developmental assembly of neural circuits. The assay exploits a classical example of heterotypic axonal interactions by modeling the sequential extension of spinal motor and somato...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2011.442
更新日期:2012-01-26 00:00:00
abstract::Genome editing harnesses programmable nucleases to cut and paste genetic information in a targeted manner in living cells and organisms. Here, I review the development of programmable nucleases, including zinc finger nucleases (ZFNs), TAL (transcription-activator-like) effector nucleases (TALENs) and CRISPR (cluster o...
journal_title:Nature protocols
pub_type: 杂志文章,评审
doi:10.1038/nprot.2016.104
更新日期:2016-09-01 00:00:00
abstract::To uncover the function of and interplay between the mammalian cytosine modifications 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC), new techniques and advances in current technology are needed. To this end, we have developed oxidative bisulfite sequencing (oxBS-seq), which can quantitatively locate 5mC an...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2013.115
更新日期:2013-10-01 00:00:00
abstract::The radial arm water maze (RAWM) contains six swim paths (arms) extending out of an open central area, with an escape platform located at the end of one arm (the goal arm). The goal arm location remains constant for a given mouse. On day 1, mice are trained for 15 trials (spaced over 3 h), with trials alternating betw...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2006.275
更新日期:2006-01-01 00:00:00
abstract::Activity-based protein profiling (ABPP) has emerged as a valuable chemical proteomics method to guide the therapeutic development of covalent drugs by assessing their on-target engagement and off-target activity. We recently used ABPP to determine the serine hydrolase interaction landscape of the experimental drug BIA...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2017.159
更新日期:2018-04-01 00:00:00
abstract::Genetically encoded fluorescent proteins (FPs) are highly utilized in cell biology research to study proteins of interest or signal processes using biosensors. To perform well in specific applications, these FPs require a multitude of tailored properties. It is for this reason that they need to be optimized by using m...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/s41596-019-0250-7
更新日期:2020-02-01 00:00:00
abstract::The combination of ChIP-seq and transcriptome analysis is a compelling approach to unravel the regulation of gene expression. Several recently published methods combine transcription factor (TF) binding and gene expression for target prediction, but few of them provide an efficient software package for the community. ...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2013.150
更新日期:2013-12-01 00:00:00
abstract::Using siRNAs to genetically manipulate immune cells is important to both basic immunological studies and therapeutic applications. However, siRNA delivery is challenging because primary immune cells are often sensitive to the delivery materials and generate immune responses. We have recently developed an amphiphilic d...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/s41596-020-00418-9
更新日期:2021-01-01 00:00:00
abstract::Isolation of monoclonal antibodies is an important technique for understanding the specificities and characteristics of antibodies that underlie the humoral immune response to a given antigen. Here we describe a technique for isolating monoclonal antibodies from human peripheral blood mononuclear cells. The protocol i...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2013.117
更新日期:2013-10-01 00:00:00
abstract::This protocol and the accompanying software program called LEVER (lineage editing and validation) enable quantitative automated analysis of phase-contrast time-lapse images of cultured neural stem cells. Images are captured at 5-min intervals over a period of 5-15 d as the cells proliferate and differentiate. LEVER au...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2011.422
更新日期:2011-11-17 00:00:00
abstract::The scototaxis (dark/light preference) protocol is a behavioral model for fish that is being validated to assess the antianxiety effects of pharmacological agents and the behavioral effects of toxic substances, and to investigate the (epi)genetic bases of anxiety-related behavior. Briefly, a fish is placed in a centra...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2009.225
更新日期:2010-02-01 00:00:00
abstract::Venous access catheters used in clinics are prone to biofilm contamination, contributing to chronic and nosocomial infections. Although several animal models for studying device-associated biofilms were previously described, only a few detailed protocols are currently available. Here we provide a protocol using totall...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2016.033
更新日期:2016-03-01 00:00:00
abstract::A procedure for the synthesis of N-succinimidyl 3-iodobenzoate labeled with any iodine isotope ([*I]SIB), which is an agent used in the radioiodination of proteins and peptides, from its tin precursor N-succinimidyl 3-(tri-n-butylstannyl)benzoate (STB) is described. Also included are protocols for the synthesis of an ...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2006.99
更新日期:2006-01-01 00:00:00
abstract::Fluorescent microspheres are commonly used to assess bone blood supply in large animals, but the technique is not widely used in smaller mammals, as traditional methods such as reference blood sampling, ventilation and catheterization are not easily applied. This protocol describes a viable alternative for measuring b...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2009.190
更新日期:2009-01-01 00:00:00
abstract::A key challenge of modern biology is to uncover the functional role of the protein entities that compose cellular proteomes. To this end, the availability of reliable three-dimensional atomic models of proteins is often crucial. This protocol presents a community-wide web-based method using RaptorX (http://raptorx.uch...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2012.085
更新日期:2012-07-19 00:00:00
abstract::Post-translational protein nitration has attracted interest owing to its involvement in cellular signaling, effects on protein function and potential as biomarker of nitroxidative stress. We describe a procedure for enriching nitropeptides for mass spectrometry (MS)-based proteomics that is a simple and reliable alter...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2014.052
更新日期:2014-04-01 00:00:00
abstract::The study of angiogenic endothelial cells (ECs) has in recent years greatly stimulated multiple fields of vascular biology research. A number of cellular models and numerous complex developmental, manipulatory and tumor animal models have been developed to study angiogenesis in vitro and in vivo. To connect the versat...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2009.96
更新日期:2009-01-01 00:00:00