Articles

The Flp double cross system a simple efficient procedure for cloning DNA fragments

While conventional cloning methods using restriction enzymes and polynucleotide ligase are adequate for most DNAs, fragments made by the polymerase chain reaction are difficult to clone because the amplifying ...

Authors: Paul D Sadowski

A fluorescent cassette-based strategy for engineering multiple domain fusion proteins

The engineering of fusion proteins has become increasingly important and most recently has formed the basis of many biosensors, protein purification systems, and classes of new drugs. Currently, most fusion pr...

Authors: Kevin Truong, Ahmad Khorchid and Mitsuhiko Ikura

Expression of self-complementary hairpin RNA under the control of the rolCpromoter confers systemic disease resistance to plum pox virus without preventing local infection

Homology-dependent selective degradation of RNA, or post-transcriptional gene silencing (PTGS), is involved in several biological phenomena, including adaptative defense mechanisms against plant viruses. Small...

Authors: Tiziana Pandolfini, Barbara Molesini, Linda Avesani, Angelo Spena and Annalisa Polverari

Oligodeoxyribonucleotide probe accessibility on a three-dimensional DNA microarray surface and the effect of hybridization time on the accuracy of expression ratios

DNA microarrays are now routinely used to monitor the transcript levels of thousands of genes simultaneously. However, the array fabrication method, hybridization conditions, and oligodeoxyribonucleotide probe...

Authors: David R Dorris, Allen Nguyen, Linn Gieser, Randall Lockner, Anna Lublinsky, Marcus Patterson, Edward Touma, Timothy J Sendera, Robert Elghanian and Abhijit Mazumder

Efficiently folding and circularly permuted variants of the Sapphire mutant of GFP

The green fluorescent protein (GFP) has been widely used in cell biology as a marker of gene expression, label of cellular structures, fusion tag or as a crucial constituent of genetically encoded biosensors. ...

Authors: Otilia Zapata-Hommer and Oliver Griesbeck

In situtransduction of target cells on solid surfaces by immobilized viral vectors

For both in vitro and in vivo gene transfer applications, recombinant viral vectors have almost always been used free in solution. Some site-specificity of the delivery of viral vectors can be achieved by applyin...

Authors: David A Hobson, Mark W Pandori and Takeshi Sano

Crude extracts of bacterially expressed dsRNA can be used to protect plants against virus infections

Double-stranded RNA (dsRNA) is a potent initiator of gene silencing in a diverse group of organisms that includes plants, Caenorhabditis elegans, Drosophila and mammals. We have previously shown and patented that...

Authors: Francisco Tenllado, Belén Martínez-García, Marisol Vargas and José Ramón Díaz-Ruíz

Retrofitting BACs with G418 resistance, luciferase, and oriP and EBNA-1 – new vectors for in vitro and in vivodelivery

Bacterial artificial chromosomes (BACs) have been used extensively for sequencing the human and mouse genomes and are thus readily available for most genes. The large size of BACs means that they can generally...

Authors: Christine Magin-Lachmann, George Kotzamanis, Leonardo D'Aiuto, Ernst Wagner and Clare Huxley

Disruption of vitellogenin gene function in adult honeybees by intra-abdominal injection of double-stranded RNA

The ability to manipulate the genetic networks underlying the physiological and behavioural repertoires of the adult honeybee worker (Apis mellifera) is likely to deepen our understanding of issues such as learni...

Authors: Gro V Amdam, Zilá LP Simões, Karina R Guidugli, Kari Norberg and Stig W Omholt

Group I aptazymes as genetic regulatory switches

Allosteric ribozymes (aptazymes) that have extraordinary activation parameters have been generated in vitro by design and selection. For example, hammerhead and ligase ribozymes that are activated by small organi...

Authors: Kristin M Thompson, Heather A Syrett, Scott M Knudsen and Andrew D Ellington

Estimating the number of integrations in transformed plants by quantitative real-time PCR

When generating transformed plants, a first step in their characterization is to obtain, for each new line, an estimate of how many copies of the transgene have been integrated in the plant genome because this...

Authors: Giovanna Mason, Paolo Provero, Anna Maria Vaira and Gian Paolo Accotto

Large scale sex typing of ostriches using DNA extracted from feathers

Ostrich (Struthio camelus) breeds have been gaining increasing significance around the world. The large-scale sex determination of chicks is an important task in the development of these breeds. To date, two PCR-...

Authors: Wilson Malagó Jr, Heitor M Franco, Euclides Matheucci Jr, Adriana Medaglia and Flavio Henrique-Silva

Genetic transformation of Vitis viniferavia organogenesis

Efficient transformation and regeneration methods are a priority for successful application of genetic engineering to vegetative propagated plants such as grape. The current methods for the production of trans...

Authors: Bruno Mezzetti, Tiziana Pandolfini, Oriano Navacchi and Lucia Landi

A comparison between two brine shrimp assays to detect in vitrocytotoxicity in marine natural products

The brine shrimp lethality assay is considered a useful tool for preliminary assessment of toxicity. It has also been suggested for screening pharmacological activities in plant extracts. However, we think tha...

Authors: José Luis Carballo, Zaira L Hernández-Inda, Pilar Pérez and María D García-Grávalos

Construction and high cytoplasmic expression of a tumoricidal single-chain antibody against hepatocellular carcinoma

Hep27 monoclonal (Hep27 Mab) is an antibody against hepatocellular carcinoma. Hep27 Mab itself can inhibit the growth of a hepatocellular carcinoma cell line (HCC-S102). We attempted to produce a single-chain ...

Authors: Duanpen Sandee, Sumalee Tungpradabkul, Manae Tsukio, Tadayuki Imanaka and Masahiro Takagi

Retrovirus-delivered siRNA

The ability of transfected synthetic small interfering (si) RNAs to suppress the expression of specific transcripts has proved a useful technique to probe gene function in mammalian cells. However, high produc...

Authors: Eric Devroe and Pamela A Silver

Quantitative assessment of the use of modified nucleoside triphosphates in expression profiling: differential effects on signal intensities and impacts on expression ratios

The power of DNA microarrays derives from their ability to monitor the expression levels of many genes in parallel. One of the limitations of such powerful analytical tools is the inability to detect certain t...

Authors: Allen Nguyen, Connie Zhao, David Dorris and Abhijit Mazumder

Signal and noise in bridging PCR

In a variant of the standard PCR reaction termed bridging, or jumping, PCR the primer-bound sequences are originally on separate template molecules. Bridging can occur if, and only if, the templates contain a ...

Authors: Shumo Liu, David S Thaler and Albert Libchaber

Characterization of cationic lipid DNA transfection complexes differing in susceptability to serum inhibition

Cationic lipid DNA complexes based on DOTAP (1,2-dioleoyl-3-(trimethyammonium) propane) and mixtures of DOTAP and cholesterol (DC) have been previously optimized for transfection efficiency in the absence of s...

Authors: Godwin Nchinda, Klaus Überla and Olaf Zschörnig

Embryonic stem cells and mice expressing different GFP variants for multiple non-invasive reporter usage within a single animal

Non-invasive autofluorescent reporters have revolutionized lineage labeling in an array of different organisms. In recent years green fluorescent protein (GFP) from the bioluminescent jellyfish Aequoria Victoria ...

Authors: Anna-Katerina Hadjantonakis, Suzanne Macmaster and Andras Nagy

Low-volume jet injection for intradermal immunization in rabbits

This study tested a low-volume (20–30 μl/20–30 μg DNA) jet injection method for intradermal delivery of a DNA vaccine. Jet injection offers the advantages of a needle-less system, low-cost, rapid preparation o...

Authors: Shuxun Ren, Minglin Li, Joanne M Smith, Louis J DeTolla and Priscilla A Furth

Protein encapsulation in liposomes: efficiency depends on interactions between protein and phospholipid bilayer.

We investigated the encapsulation mechanism of enzymes into liposomes. The existing protocols to achieve high encapsulation efficiencies are basically optimized for chemically stable molecules. Enzymes, howeve...

Authors: Jacques-Philippe Colletier, Barnabé Chaize, Mathias Winterhalter and Didier Fournier

A new device for measurement of fibrin clot lysis: application to the Euglobulin Clot Lysis Time

Determination of clot lysis times on whole blood, diluted whole blood, plasma or plasma fraction has been used for many years to assess the overall activity of the fibrinolytic system. We designed a completely...

Authors: K Zouaoui Boudjeltia, Ph Cauchie, Cl Remacle, M Guillaume, D Brohée, JL Hubert and M Vanhaeverbeek

Selective Permeation and Organic Extraction of Recombinant Green Fluorescent Protein (gfp _uv ) from Escherichia coli

Transformed cells of Escherichia coli DH5-α with pGFPuv, induced by IPTG (isopropyl-β-d-thiogalactopyranoside), express the green fluorescent protein (gfp _uv ) during growth ...

Authors: Thereza Christina Vessoni Penna and Marina Ishii

Glycerol restores heat-induced p53-dependent apoptosis of human glioblastoma cells bearing mutant p53

We have previously reported that glycerol acts as a chemical chaperone to restore the expression of WAF1 in some human cancer cell lines bearing mutant p53. Since the expression of WAF1 is up-regulated by acti...

Authors: Takeo Ohnishi, Ken Ohnishi and Akihisa Takahashi

Effective generation of transgenic pigs and mice by linker based sperm-mediated gene transfer.

Transgenic animals have become valuable tools for both research and applied purposes. The current method of gene transfer, microinjection, which is widely used in transgenic mouse production, has only had limi...

Authors: Keejong Chang, Jin Qian, MeiSheng Jiang, Yi-Hsin Liu, Ming-Che Wu, Chi-Dar Chen, Chao-Kuen Lai, Hsin-Lung Lo, Chin-Ton Hsiao, Lucy Brown, James Bolen Jr, Hsiao-I Huang, Pei-Yu Ho, Ping Yao Shih, Chen-Wen Yao, Wey-Jinq Lin…

Genetically modified parthenocarpic eggplants: improved fruit productivity under both greenhouse and open field cultivation.

Parthenocarpy, or fruit development in the absence of fertilization, has been genetically engineered in eggplant and in other horticultural species by using the DefH9-iaaM gene. The iaaM gene codes for tryptophan...

Authors: Nazzareno Acciarri, Federico Restaino, Gabriele Vitelli, Domenico Perrone, Michela Zottini, Tiziana Pandolfini, Angelo Spena and Giuseppe Leonardo Rotino

Rhodococcus erythropolis ATCC 25544 as a suitable source of cholesterol oxidase: cell-linked and extracellular enzyme synthesis, purification and concentration

The suitability of the strain Rhodococcus erythropolis ATCC 25544 grown in a two-liter fermentor as a source of cholesterol oxidase has been investigated. The strain produces both cell-linked and extracellular ch...

Authors: Mar M Sojo, Roque R Bru and Francisco F García-Carmona

Expression of a crown gall biological control phenotype in an avirulent strain of Agrobacterium vitis by addition of the trifolitoxin production and resistance genes

Agrobacterium vitis is a causal agent of crown-gall disease. Trifolitoxin (TFX) is a peptide antibiotic active only against members of a specific group of α-proteobacteria that includes Agrobacterium and its clos...

Authors: Thomas C Herlache and Eric W Triplett

Optimisation of transgene action at the post-transcriptional level: high quality parthenocarpic fruits in industrial tomatoes

Genetic engineering of parthenocarpy confers to horticultural plants the ability to produce fruits under environmental conditions that curtail fruit productivity and quality. The DefH9-iaaM transgene, whose predi...

Authors: Tiziana Pandolfini, Giuseppe L Rotino, Serena Camerini, Roberto Defez and Angelo Spena

A combined in vitro / in vivo selection for polymerases with novel promoter specificities

The DNA-dependent RNA polymerase from T7 bacteriophage (T7 RNAP) has been extensively characterized, and like other phage RNA polymerases it is highly specific for its promoter. A combined in vitro / in vivo sele...

Authors: Jijumon Chelliserrykattil, George Cai and Andrew D Ellington

Generation of single-copy transgenic mouse embryos directly from ES cells by tetraploid embryo complementation

Transgenic mice have been used extensively to analyze gene function. Unfortunately, traditional transgenic procedures have only limited use in analyzing alleles that cause lethality because lines of founder mi...

Authors: Ravi P Misra, Sarah K Bronson, Qi Xiao, Wendy Garrison, Jixuan Li, Roong Zhao and Stephen A Duncan

Manipulation of gene expression by an ecdysone-inducible gene switch in tumor xenografts

Rapid, robust and reversible induction of transgene expression would significantly facilitate cancer gene therapy as well as allow the in vivo functional study of newly discovered genes in tumor formation and ...

Authors: Larry R Karns PhD, Anne Kisielewski BSc, Kathryn M Gulding BSc and Dan Theodorescu MD PhD

Long term adaptation of a microbial population to a permanent metabolic constraint: overcoming thymineless death by experimental evolution of Escherichia coli

To maintain populations of microbial cells under controlled conditions of growth and environment for an indefinite duration is a prerequisite for experimentally evolving natural isolates of wild-type species o...

Authors: Valérie A de Crécy-Lagard, Jacques Bellalou, Rupert Mutzel and Philippe Marlière

L-Glutamate production by lysozyme-sensitive Corynebacterium glutamicum ltsAmutant strains

A non-pathogenic species of coryneform bacteria, Corynebacterium glutamicum, was originally isolated as an L-glutamate producing bacterium and is now used for fermentative production of various amino acids. A mut...

Authors: Takashi Hirasawa, Masaaki Wachi and Kazuo Nagai

A simple method for statistical analysis of intensity differences in microarray-derived gene expression data

Microarray experiments offer a potent solution to the problem of making and comparing large numbers of gene expression measurements either in different cell types or in the same cell type under different condi...

Authors: Alexander Kamb and Mani Ramaswami

Mutant loxP vectors for selectable marker recycle and conditional knock-outs

Gene disruption by targeted integration of transfected constructs becomes increasingly popular for studies of gene function. The chicken B cell line DT40 has been widely used as a model for gene knock-outs due...

Authors: Hiroshi Arakawa, Dimitry Lodygin and Jean-Marie Buerstedde

A surrogate-based approach for post-genomic partner identification

Modern drug discovery is concerned with identification and validation of novel protein targets from among the 30,000 genes or more postulated to be present in the human genome. While protein-protein interactio...

Authors: Renuka C Pillutla, Ku-chuan Hsiao, Renee Brissette, Paul S Eder, Tony Giordano, Paul W Fletcher, Michael Lennick, Arthur J Blume and Neil I Goldstein

Mannosylerythritol lipid, a yeast extracellular glycolipid, shows high binding affinity towards human immunoglobulin G

There have been many attempts to develop new materials with stability and high affinity towards immunoglobulins. Some of glycolipids such as gangliosides exhibit a high affinity toward immunoglobulins. However...

Authors: Jae Hong Im, Takashi Nakane, Hiroshi Yanagishita, Toru Ikegami and Dai Kitamoto

Oriented antibody immobilization to polystyrene macrocarriers for immunoassay modified with hydrazide derivatives of poly(meth)acrylic acid

Hydrophobic polystyrene is the most common material for solid phase immunoassay. Proteins are immobilized on polystyrene by passive adsorption, which often causes considerable denaturation. Biological macromol...

Authors: Vadim V Shmanai, Tamara A Nikolayeva, Ludmila G Vinokurova and Anatoli A Litoshka

Erratum to: The evaluation of yeast derivatives as adjuvant of the immune response to the Bm86 antigen in cattle: Correction

Authors: Manuel Rodríguez Valle, Carlos Montero, Héctor Machado and Marisdania Joglar

The evaluation of yeast derivatives as adjuvants for the immune response to the Bm86 antigen in cattle

The Gavac™ vaccine against the cattle tick Boophilus microplus has proven its efficacy in a large number of controlled and field experiments. However, this vaccine could be further improved by searching for new a...

Authors: Manuel Rodríguez Valle, Carlos Montero, Héctor Machado and Marisdania Joglar

Inhibition of spontaneous induction of lambdoid prophages in Escherichia colicultures: simple procedures with possible biotechnological applications

Infections of bacterial cultures by bacteriophages are serious problems in biotechnological laboratories. Apart from such infections, prophage induction in the host cells may also be dangerous. Escherichia coli i...

Authors: Agata Czyz, Marcin Los, Borys Wrobel and Grzegorz Wegrzyn