Cloning of a novel omega-6 desaturase from flax (Linum usitatissimum L.) and its functional analysis in Saccharomyces cerevisiae.

Abstract:

:The Delta12 desaturase represents a diverse gene family in plants and is responsible for conversion of oleic acid (18:1) to linoleic acid (18:2). Several members of this family are known from plants like Arabidopsis and Soybean. Using primers from conserved C- and N-terminal regions, we have cloned a novel Delta12 desaturase gene amplified from flax genomic DNA, denoted as LuFAD2-2. This intron-less gene is 1,149-base pair long encoding 382 amino acids-putative membrane-bound Delta12 desaturase protein. Sequence comparisons show that the novel sequence has 85% similarity with previously reported flax Delta12 desaturase at amino acid level and shows typical features of membrane-bound desaturase such as three conserved histidine boxes along with four membrane-spanning regions that are universally present among plant desaturases. The signature amino acid sequence 'YNNKL' was also found to be present at the N terminus of the protein, which is necessary and sufficient for ER localization of enzyme. Neighbor-Joining tree generated from the sequence alignment grouped LuFAD2-2 among the other FAD2 sequences from Ricinus, Hevea, Jatropha, and Vernicia. When LuFAD2-2 and LuFAD2 were expressed in Saccharomyces cerevisiae, they could convert the oleic acid to linoleic acid, with an average conversion rate of 5.25 and 8.85%, respectively. However, exogenously supplied linoleic acid was feebly converted to linolenic acid suggesting that LuFAD2-2 encodes a functional FAD2 enzyme and has substrate specificity similar to LuFAD2.

journal_name

Mol Biotechnol

journal_title

Molecular biotechnology

authors

Khadake RM,Ranjekar PK,Harsulkar AM

doi

10.1007/s12033-009-9150-3

subject

Has Abstract

pub_date

2009-06-01 00:00:00

pages

168-74

issue

2

eissn

1073-6085

issn

1559-0305

journal_volume

42

pub_type

杂志文章
  • Deciphering enzyme function using peptide arrays.

    abstract::Enzymes are key molecules in signal-transduction pathways. However, only a small fraction of more than 500 human kinases, 300 human proteases and 200 human phosphatases is characterised so far. Peptide microarray based technologies for extremely efficient profiling of enzyme substrate specificity emerged in the last y...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章,评审

    doi:10.1007/s12033-011-9402-x

    authors: Thiele A,Stangl GI,Schutkowski M

    更新日期:2011-11-01 00:00:00

  • Immunoglobulin fusion proteins as a tool for evaluation of T-cell costimulatory molecules.

    abstract::Costimulatory signal(s) in addition to engagement of T cell receptor is important for optimal activation of T lymphocytes. During last decade several dozens of new costimulatory molecules, including 4-1BB, CD40, OX40, CD27, and ICOS, have been identified. It is likely that in Post-Genome Era more molecules with potent...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1385/MB:21:3:259

    authors: Chapoval AI,Zhu G,Chen L

    更新日期:2002-07-01 00:00:00

  • Transplantation of mesenchymal cells improves peripheral limb ischemia in diabetic rats.

    abstract::Adipose tissue-derived mesenchymal stromal cells (ADSCs) are a prominent cellular source for regenerative medicine. We tested whether transplantation of ADSCs into the ischemic muscular tissue of diabetic animals would attenuate impaired cell metabolism and microcirculatory function. We induced unilateral hind limb is...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-014-9735-3

    authors: Madonna R,Delli Pizzi S,Tartaro A,De Caterina R

    更新日期:2014-05-01 00:00:00

  • MALDI-TOF mass spectrometry: a versatile tool for high-performance DNA analysis.

    abstract::Matrix-assisted laser desorption/ionization (MALDI) time-of-flight (TOF) mass spectrometry (MS) has developed during the past decade into a versatile tool for biopolymer analysis. The aim of this review is to summarize this development and outline the applications, which have been enabled for routine use in the field ...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章,评审

    doi:10.1385/MB:26:2:147

    authors: Jurinke C,Oeth P,van den Boom D

    更新日期:2004-02-01 00:00:00

  • Optimisation of the Production and Bleaching Process for a New Laccase from Madurella mycetomatis, Expressed in Pichia pastoris: from Secretion to Yielding Prominent.

    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

    authors: Tülek A,Karataş E,Çakar MM,Aydın D,Yılmazcan Ö,Binay B

    更新日期:2021-01-01 00:00:00

  • Isolation of the 5'-end of plant genes from genomic DNA by TATA-box-based degenerate primers.

    abstract::We report a rapid and simple method for isolating the 5'-end of plant genes from genomic DNA by polymerase chain reaction (PCR) with TATA-box-based degenerate primers (TDPs). The TDPs were specially designed according to the TATA box, which is conserved in the promoter region of most plant genes. The unknown 5'-ends o...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-010-9323-0

    authors: Guo Y,Ma L,Ji Y,Pu G,Liu B,Du Z,Li G,Ye H,Wang H

    更新日期:2011-02-01 00:00:00

  • Early changes in gene expression induced by acute UV exposure in leaves of Psychotria brachyceras, a bioactive alkaloid accumulating plant.

    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

    authors: do Nascimento NC,Menguer PK,Sperotto RA,de Almeida MR,Fett-Neto AG

    更新日期:2013-05-01 00:00:00

  • High-level expression of heme-dependent catalase gene katA from Lactobacillus Sakei protects Lactobacillus rhamnosus from oxidative stress.

    abstract::Lactic acid bacteria (LAB) are generally sensitive to hydrogen peroxide (H(2)O(2)), Lactobacillus sakei YSI8 is one of the very few LAB strains able to degrade H(2)O(2) through the action of a heme-dependent catalase. Lactobacillus rhamnosus strains are very important probiotic starter cultures in meat product ferment...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-010-9254-9

    authors: An H,Zhou H,Huang Y,Wang G,Luan C,Mou J,Luo Y,Hao Y

    更新日期:2010-06-01 00:00:00

  • Cloning, sequence and functional analysis of goat ATP-binding cassette transporter G2 (ABCG2).

    abstract::ATP-binding cassette transporter G2 (ABCG2) gene encodes a protein that has a wide variety of substrates and is responsible for the active secretion of clinically and toxicologically important molecules into milk. Although known in many species, this marks the first time this gene product has been reported in goats. I...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-007-9024-5

    authors: Wu HJ,Luo J,Wu N,Matand K,Zhang LJ,Han XF,Yang BJ

    更新日期:2008-05-01 00:00:00

  • Gene transfer by electroporation.

    abstract::Electroporation of cells in the presence of DNA is widely used for the introduction of transgenes either stably or transiently into bacterial, fungal, animal, and plant cells. A review of the literature shows that electroporation parameters are often reported in an incomplete or incorrect manner, forcing researchers t...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章,评审

    doi:10.1007/BF02821542

    authors: Lurquin PF

    更新日期:1997-02-01 00:00:00

  • Rapid and efficient gene splicing using megaprimer-based protocol.

    abstract::Megaprimer-based methodology has been widely applied in site-directed mutagenesis, but rarely used in gene splicing. In this article, we describe a modification of the megaprimer PCR method, which can efficiently create and amplify a specific ligated chimeric gene segment in a PCR reaction and under a common PCR progr...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-008-9078-z

    authors: Chen JR,Lü JJ,Wang HF

    更新日期:2008-11-01 00:00:00

  • High Conversion of D-Fructose into D-Allulose by Enzymes Coupling with an ATP Regeneration System.

    abstract::D-Allulose is a rare monosaccharide that exists in extremely small quantities in nature, and it is also hard to prepare at a large scale via chemical or enzyme synthetic route due to low conversion and downstream separation complexity. Using D-psicose epimerase and L-rhamnulose kinase, a method enabling high conversio...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-019-00174-6

    authors: Xiao Q,Niu J,Liu H,Liu Y,Zhou X

    更新日期:2019-06-01 00:00:00

  • Expression of Recombinant PfCelTOS Antigen in the Chloroplast of Chlamydomonas reinhardtii and its Potential Use in Detection of Malaria.

    abstract::Malaria is a serious but preventable and treatable infectious disease that is found in over 100 countries around the world. Correct and rapid diagnosis of malaria infection can rescue the patient of getting sicker and reduces the risk of disease spreading among humans. Chlamydomonas reinhardtii chloroplast is an attra...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-018-0140-1

    authors: Shamriz S,Ofoghi H

    更新日期:2019-02-01 00:00:00

  • Toward more efficient protein expression: keep the message simple.

    abstract::Optimization of gene coding-sequence, including preferred codon usage and removal of cryptic splice sites and mRNA-destabilizing motifs, has been shown to improve recombinant protein production of different proteins. Here, we present data to show that gene optimization can also be used to improve the production of a c...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1385/MB:34:2:151

    authors: Kalwy S,Rance J,Young R

    更新日期:2006-10-01 00:00:00

  • Oligovalent Fab display on M13 phage improved by directed evolution.

    abstract::Efficient display of antibody on filamentous phage M13 coat is crucial for successful biopanning selections. We applied a directed evolution strategy to improve the oligovalent display of a poorly behaving Fab fragment fused to phage gene-3 for minor coat protein (g3p). The Fab displaying clones were enriched from a r...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-009-9231-3

    authors: Huovinen T,Sanmark H,Ylä-Pelto J,Vehniäinen M,Lamminmäki U

    更新日期:2010-03-01 00:00:00

  • Virus crystallography.

    abstract::Virus crystallography can provide atomic resolution structures for intact isometric virus particles and components thereof. The methodology is illustrated by reference to a particularly complex example, the core of the bluetongue virus (700 A). ...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章,评审

    doi:10.1385/MB:12:1:13

    authors: Fry EE,Grimes J,Stuart DI

    更新日期:1999-08-01 00:00:00

  • A stable human-cell system overexpressing cystic fibrosis transmembrane conductance regulator recombinant protein at the cell surface.

    abstract::Recent human clinical trials results demonstrated successful treatment for certain genetic forms of cystic fibrosis (CF). To extend treatment opportunities to those afflicted with other genetic forms of CF disease, structural and biophysical characterization of CF transmembrane conductance regulator (CFTR) is urgently...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-014-9830-5

    authors: Hildebrandt E,Mulky A,Ding H,Dai Q,Aleksandrov AA,Bajrami B,Diego PA,Wu X,Ray M,Naren AP,Riordan JR,Yao X,DeLucas LJ,Urbatsch IL,Kappes JC

    更新日期:2015-05-01 00:00:00

  • Reactivation of transgene expression by alleviating CpG methylation of the Rous sarcoma virus promoter in transgenic quail cells.

    abstract::In this study, we investigated the relative expression of the Rous sarcoma virus (RSV) promoter-driven expression of enhanced green fluorescent protein (EGFP) in fibroblasts of transgenic quails. We analyzed the direct influence of CpG methylation of the RSV promoter on the transcriptional activity of delivered transg...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-011-9393-7

    authors: Jang HJ,Choi JW,Kim YM,Shin SS,Lee K,Han JY

    更新日期:2011-11-01 00:00:00

  • Improved method for generating microarray probes using submicrogram amounts of total RNA.

    abstract::The ability to identify disease-associated genes using microarray technology is dependent on isolating high-quality total RNA from the diseased tissue under study. However, it is not always possible to obtain large amounts of affected tissue from patients and therefore the quality and yield of RNA may be compromised. ...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1385/MB:23:2:171

    authors: Murray EJ

    更新日期:2003-02-01 00:00:00

  • Construction and characterization of a bacterial artificial chromosome library of the causal agent of Black Sigatoka fungal leaf spot disease of banana and plantain, Mycosphaerella fijiensis.

    abstract::A bacterial artificial chromosome library of the causal agent of the Black Sigatoka leaf spot disease of banana and plantain, Mycosphaerella fijiensis, has been constructed using a non-sphaeroplasting technique and characterized using both homologous and heterologous probes. After first and a second size selection of ...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-007-0016-2

    authors: Canto-Canché B,Guillén-Maldonado DK,Peraza-Echeverría L,Conde-Ferráez L,James-Kay A

    更新日期:2007-05-01 00:00:00

  • Expression of human rotavirus chimeric fusion proteins from replicating but non disseminating adenovectors and elicitation of rotavirus-specific immune responses in mice.

    abstract::The aim of this study was to evaluate the usefulness of replicating but non disseminating adenovirus vectors (AdVs) as vaccine vector using human rotavirus (HRV) as a model pathogen. HRV VP7, VP4, or VP4Δ (N-terminal 336 amino acids of VP4) structural proteins as well as the VP4Δ::VP7 chimeric fusion protein were expr...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-013-9653-9

    authors: Girard A,Roques E,St-Louis MC,Massie B,Archambault D

    更新日期:2013-07-01 00:00:00

  • Molecular factors affecting the accumulation of recombinant proteins in the Chlamydomonas reinhardtii chloroplast.

    abstract::In an effort to develop microalgae as a robust system for the production of valuable proteins, we analyzed some of the factors affecting recombinant protein expression in the chloroplast of the green alga Chlamydomonas reinhardtii. We monitored mRNA accumulation, protein synthesis, and protein turnover for three codon...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-010-9348-4

    authors: Coragliotti AT,Beligni MV,Franklin SE,Mayfield SP

    更新日期:2011-05-01 00:00:00

  • A flow cytometric protocol for titering recombinant adenoviral vectors containing the green fluorescent protein.

    abstract::As the use of adenoviral vectors in gene therapy protocols increases, there is a corresponding need for rapid, accurate, and reproducible titer methods. Multiple methods currently exist for determining titers of recombinant adenoviral vector, including optical absorbance, electron microscopy, fluorescent focus assay, ...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1385/MB:14:3:197

    authors: Hitt DC,Booth JL,Dandapani V,Pennington LR,Gimble JM,Metcalf J

    更新日期:2000-03-01 00:00:00

  • Expression of modified Cry1Ac gene of Bacillus thuringiensis in transgenic tobacco plants.

    abstract::Several mutations were introduced into the Cry1Ac toxin gene, resulting in four variants with altered sequences that were responsible for low expression of the toxin in transgenic plants. These variants were as follows: V1, with modified three A/T-rich regions, including the first signal of transcription termination; ...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1385/mb:26:1:17

    authors: Misztal LH,Mostowska A,Skibinska M,Bajsa J,Musial WG,Jarmolowski A

    更新日期:2004-01-01 00:00:00

  • In vitro direct repeats-mediated deletion during PCR amplification.

    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

    authors: Hadj Kacem B,Gargouri J,Gargouri A

    更新日期:2008-09-01 00:00:00

  • Genomic imprinting and chromatin insulation in Beckwith-Wiedemann syndrome.

    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

    authors: Greally JM

    更新日期:1999-04-01 00:00:00

  • Carotenoids and flavonoids contribute to nutritional protection against skin damage from sunlight.

    abstract::The concept of photoprotection by dietary means is gaining momentum. Plant constituents such as carotenoids and flavonoids are involved in protection against excess light in plants and contribute to the prevention of UV damage in humans. As micronutrients, they are ingested with the diet and are distributed into light...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章,评审

    doi:10.1007/s12033-007-0051-z

    authors: Stahl W,Sies H

    更新日期:2007-09-01 00:00:00

  • Analysis of protein glycosylation by mass spectrometry.

    abstract::There is a growing pharmaceutical market for protein-based drugs for use in therapy and diagnosis. The rapid developments in molecular and cell biology have resulted in production of expression systems for manufacturing of recombinant proteins and monoclonal antibodies. These proteins are glycosylated when expressed i...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章,评审

    doi:10.1007/BF02745880

    authors: Nilsson B

    更新日期:1994-12-01 00:00:00

  • Analysis of HOX gene expression patterns in human breast cancer.

    abstract::HOX genes are highly conserved transcription factors that determine the identity of cells and tissues along the anterior-posterior body axis in developing embryos. Aberrations in HOX gene expression have been shown in various tumors. However, the correlation of HOX gene expression patterns with tumorigenesis and cance...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-013-9682-4

    authors: Hur H,Lee JY,Yun HJ,Park BW,Kim MH

    更新日期:2014-01-01 00:00:00

  • Development of an efficient protocol of RNA isolation from recalcitrant tree tissues.

    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 utiliz...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-007-0073-6

    authors: Wang X,Tian W,Li Y

    更新日期:2008-01-01 00:00:00