Abstract：
:Isolation of RNA from recalcitrant tree tissues has been problematic due to large amounts of secondary metabolites and interfering compounds in their cells. We have developed an efficient RNA extraction method, which yielded high-quality RNA preparations from tissues of the lychee tree. The method reported here utilized EDTA, LSS, and CTAB to successfully inhibit RNase activities. It was found that a high ionic strength brought about by 2 M NaCl was necessary. In addition, secondary metabolites and other interfering compounds were effectively removed using sodium borate and PVPP under a deoxidized condition. The quality of purified RNA was tested by both RACE and Northern blotting analysis, ensuring that the RNA could be used for subsequent gene expression analysis. This method has been successfully applied to purify RNA from 15 other plant species. In conclusion, the protocol reported here is expected to have excellent applications for RNA isolation from recalcitrant plant tissues.
journal_name
Mol Biotechnoljournal_title
Molecular biotechnologyauthors
Wang X,Tian W,Li Ydoi
10.1007/s12033-007-0073-6subject
Has Abstractpub_date
2008-01-01 00:00:00pages
57-64issue
1eissn
1073-6085issn
1559-0305journal_volume
38pub_type
杂志文章abstract：:A functional assay for proteolytic processing of the amyloid precursor protein (APP) was set up in yeast. This consisted of a membrane-bound chimeric protein containing the beta-secretase cleaved C-terminal fragment of APP fused to the Ga14 transcription factor. Using this chimera in a GAL-reporter yeast strain, an ex...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1385/MB:15:1:29
更新日期：2000-05-01 00:00:00
abstract：:Neuroblastoma is the most common extracranial solid tumor of infancy. Amplification of MYCN oncogene is found in approximately 20 % of all neuroblastoma patients and correlates with advanced disease stages, rapid tumor progression, and poor prognosis, making this gene an obvious therapeutic target. However, being a tr...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-014-9739-z
更新日期：2014-07-01 00:00:00
abstract：:In this study, human alpha-1,4-N-acetylglucosaminyltransferase (alpha4GnT) fused with GFP(uv) (GFP(uv)-alpha4GnT) was expressed using both a transformed cell system and silkworm larvae. A Tn-pXgp-GFP(uv)-alpha4GnT cell line, isolated after expression vector transfection, produced 106 mU/ml of alpha4GnT activity in sus...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-009-9174-8
更新日期：2009-09-01 00:00:00
abstract：:Pasteurella multocida is the main cause of haemorrhagic septicaemia (HS) outbreak in livestock, such as cattle and buffaloes. Conventional vaccines such as alum-precipitated or oil-adjuvant broth bacterins were injected subcutaneously to provide protection against HS. However, the immunity developed is only for short ...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-020-00244-0
更新日期：2020-05-01 00:00:00
abstract：:Genes are recognized as undergoing genomic imprinting when they are capable of being expressed only from the paternal or only from the maternal chromosome. The process can occur coordinately within large physical domains in mammalian chromosomes. One interesting facet of the study of genomic imprinting is that it offe...
journal_title：Molecular biotechnology
pub_type： 杂志文章,评审
doi：10.1007/BF02915809
更新日期：1999-04-01 00:00:00
abstract：:IMGT, the International ImMunoGeneTics information system ( http://imgt.cines.fr ), was created in 1989 by the Laboratoire d'ImmunoGénétique Moléculaire (LIGM) (Université Montpellier 2 and CNRS) at Montpellier, France, in order to standardize and manage the complexity of immunogenetics data. IMGT is recognized as the...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-008-9062-7
更新日期：2008-09-01 00:00:00
abstract：:Recombinant proteins are useful tools in biological research, drug development, and drug screening. Specially designed expression vectors have been developed to introduce cDNA for recombinant protein expression in mammalian cells. We have combined a dicistronic mRNA design for expression of the recombinant protein, us...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/BF02745860
更新日期：1998-08-01 00:00:00
abstract：:Isolation of high quality nucleic acids from plant tissues rich in polysaccharides and polyphenols is often difficult. The presence of these substances can affect the quality and/or quantity of the nucleic acids isolated. Here, we describe a rapid and efficient nucleic acids extraction protocol that in contrast to oth...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-011-9384-8
更新日期：2011-10-01 00:00:00
abstract：:Water-soluble quinoprotein glucose dehydrogenase (PQQGDH-B) is a dimeric enzyme whose application for glucose sensing is the focus of much attention. We attempted to increase the thermal stability of PQQGDH-B by introducing a disulfide bond at the dimer interface. The Ser residue at position 415 was selected for subst...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1385/MB:24:2:97
更新日期：2003-06-01 00:00:00
abstract：:Human adipose tissue-derived stromal cells (ADSCs) are being evaluated for cardiovascular repair. We developed an ex vivo method for producing angiogenic ADSCs transduced with a self-inactivating lentiviral vector (LV) expressing the enhanced green fluorescence protein (EGFP) from an internal cytomegalovirus (CMV) pro...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-012-9537-4
更新日期：2013-05-01 00:00:00
abstract：:The biosensor field has grown enormously since the first demonstration of the biosensor concept by Leland C. Clark, Jr. in 1962. Today's biosensor market is dominated by glucose biosensors, mass-produced enzyme electrodes for the rapid self-diagnosis of blood glucose levels by diabetes sufferers. Here we take a histor...
journal_title：Molecular biotechnology
pub_type： 杂志文章,评审
doi：10.1385/MB:32:3:249
更新日期：2006-03-01 00:00:00
abstract：:Since it was first introduced in late 1990s Wave bioreactor has been used for protein production by mammalian and insect cell lines. However, using Wave bioreactor to produce human monoclonal antibody by stable Drosophila Schneider 2 (S2) cell transfectants has not been reported before. In this study, S2 cells were co...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-011-9484-5
更新日期：2012-10-01 00:00:00
abstract：:The macrofungus Cordyceps militaris contains many kinds of bioactive ingredients that are regulated by functional genes, but the functions of many genes in C. militaris are still unknown. In this study, to improve the frequency of homologous integration, a genetic transformation system based on a split-marker approach...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-018-0080-9
更新日期：2018-05-01 00:00:00
abstract：:The monomeric bacteriophage RNA polymerases allow the synthesis of virtually any RNA molecule in unlimited quantity. In this protocol, we describe the preparation of plasmid and PCR-derived templates. A basic transcription protocol is provided with several optional modifications. The use of RNA probes in Northern blot...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/BF02761751
更新日期：1997-04-01 00:00:00
abstract：:Tea, a beverage crop, is a rich source of polyphenols and polysaccharides which greatly attribute to its importance. However, oxidation and precipitation of these compounds during nucleic acids extraction is a limitation to molecular biology and genomic studies. On isolation of total RNA from root tissue using establi...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-011-9476-5
更新日期：2012-09-01 00:00:00
abstract：:This article presents a simple and rapid method for removal of unincorporated label and proteins from DNA sequencing reactions by using Wizard purification resin. This method can be successfully applied for preparation of end-labeled oligonucleotides free of unincorporated label, which is important in experiments (inc...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/BF02789066
更新日期：1996-04-01 00:00:00
abstract：:Laccases are polyphenol oxidoreductases used in a number of industrial applications. Due to the increasing demand for these "green catalysis" enzymes, the identification and biochemical characterisation of their novel properties is essential. In our study, cloned Madurella mycetomatis laccase (mmlac) genes were hetero...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-020-00281-9
更新日期：2021-01-01 00:00:00
abstract：:While conducting our research on mutations in the human blood platelet glycoprotein Ib-alpha (GPIbalpha) gene, we detected an unusual deletion of 84 bp. This deletion took place in vitro, during PCR and between two direct repeats. It was observed that the deletion could be detected either by the direct sequencing of t...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-008-9059-2
更新日期：2008-09-01 00:00:00
abstract：:De novo design provides an in silico toolkit for the design of novel small molecular structures to a set of specified structural constraints. With the avalanche of bioinformatics data, de novo design is ideally suited for exploring molecules that could be useful for chemical genomics. The design process involves manip...
journal_title：Molecular biotechnology
pub_type： 杂志文章,评审
doi：10.1007/s12033-007-0037-x
更新日期：2007-11-01 00:00:00
abstract：:In mammals, CpG methylation is one of the mechanisms of epigenetic control over the linear sequence of bases of deoxyribonucleic acid (DNA); about 70% of CpG dinucleotides are methylated. The actual signal that triggers DNA methylation is not known, although repetitive DNA has been shown to be an attractive template f...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1385/MB:26:3:215
更新日期：2004-03-01 00:00:00
abstract：:UV-B radiation can damage biomolecules, such as DNA, RNA, and proteins, halting essential cellular processes; this damage is partly due to ROS generation. Plant secondary metabolites may protect against UV-B. Psychotria brachyceras Müll. Arg. (Rubiaceae), a subtropical shrub, produces brachycerine, a monoterpene indol...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-012-9546-3
更新日期：2013-05-01 00:00:00
abstract：:Technological achievements in the last 5 to 10 yr and their application to sequencing and polymorphism discovery in the human genome have fostered a renewed interest in the genetic basis of drug response. Consequently, the field of pharmacogenetics/pharmacogenomics has been gaining momentum, fueled not only on technol...
journal_title：Molecular biotechnology
pub_type： 杂志文章,评审
doi：10.1385/MB:25:3:275
更新日期：2003-11-01 00:00:00
abstract：:Classic toxicology studies often utilize in vivo animal models. Newer approaches employing in vitro organ-specific cellular models have been developed in recent years to help accelerate the speed and reduce the cost of traditional toxicology testing. Toward the goal of supporting in vitro cellular model research with ...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-020-00245-z
更新日期：2020-04-01 00:00:00
abstract：:Coptotermes curvignathus is a termite that, owing to its ability to digest living trees, serves as a gold mine for robust industrial enzymes. This unique characteristic reflects the presence of very efficient hydrolytic enzyme systems including cellulases. Transcriptomic analyses of the gut of C. curvignathus revealed...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-017-0015-x
更新日期：2017-07-01 00:00:00
abstract：:Translation initiation of Coxsackievirus B3 (CVB3) RNA is directed by an internal ribosome entry site (IRES) within the 5' untranslated region. Host cell factors involved in this process include some canonical translation factors and additional RNA-binding proteins. We have, previously, described that the Sabin3-like ...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-012-9592-x
更新日期：2013-06-01 00:00:00
abstract：:Many preclinical studies have shown RNA interference (RNAi) as a new promising way to treat various human diseases including cancer and virus infection and there is an increasing demand for the large-scale preparation of short interfering RNAs (siRNAs) at low cost. Data are accumulating to show that endoribonuclease-p...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1385/MB:31:3:203
更新日期：2005-11-01 00:00:00
abstract：:We have modified the differential display of 3'-end restriction fragments of cDNA technique by combining it with the amplified fragment length polymorphism (AFLP) approach and silver staining. Modified oligo-dT primers were used for a reverse transcription step. ds cDNA was digested with the Mse I restriction enzyme a...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1385/MB:12:2:137
更新日期：1999-09-01 00:00:00
abstract：:Small ubiquitin-related modifier (SUMO) fusion system has been shown to be efficient for enhancing expression and preventing degradation of the target protein. We showed herein that SUMO fusion to human keratinocyte growth factor 2 (hKGF-2) gene was feasible and it significantly enhanced protein expression and its eff...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-008-9135-7
更新日期：2009-05-01 00:00:00
abstract：:The Embrapa 5.1 genetically modified (GM) common bean was approved for commercialization in Brazil. Methods for the quantification of this new genetically modified organism (GMO) are necessary. The development of a suitable endogenous reference is essential for GMO quantification by real-time PCR. Based on this, a new...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-014-9786-5
更新日期：2014-11-01 00:00:00
abstract：:Bacteriophage lambda has been in use as a cloning vector for over 25 years, and has been used extensively as an expression vector. The efficiency of packaging and infection, and the simplicity of plaque screening are advantages of lambda as a cloning vector. A number of ingenious modifications help overcome the disadv...
journal_title：Molecular biotechnology
pub_type： 杂志文章,评审
doi：10.1385/MB:17:3:219
更新日期：2001-03-01 00:00:00
