Targeted Gene Deletion in Cordyceps militaris Using the Split-Marker Approach.

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 was developed for the first time in C. militaris to knock out a gene encoding a terpenoid synthase (Tns). The linear and split-marker deletion cassettes were constructed and introduced into C. militaris protoplasts by PEG-mediated transformation. The transformation of split-marker fragments resulted in a higher efficiency of targeted gene disruption than the transformation of linear deletion cassettes did. The color phenotype of the Tns gene deletion mutants was different from that of wild-type C. militaris. Moreover, a PEG-mediated protoplast transformation system was established, and stable genetic transformants were obtained. This method of targeted gene deletion represents an important tool for investigating the role of C. militaris genes.

journal_name

Mol Biotechnol

journal_title

Molecular biotechnology

authors

Lou H,Ye Z,Yun F,Lin J,Guo L,Chen B,Mu Z

doi

10.1007/s12033-018-0080-9

subject

Has Abstract

pub_date

2018-05-01 00:00:00

pages

380-385

issue

5

eissn

1073-6085

issn

1559-0305

pii

10.1007/s12033-018-0080-9

journal_volume

60

pub_type

杂志文章
  • IMGT, the International ImMunoGeneTics Information System for Immunoinformatics : methods for querying IMGT databases, tools, and web resources in the context of immunoinformatics.

    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

    authors: Lefranc MP

    更新日期:2008-09-01 00:00:00

  • Biochemical and Molecular Characterizations of a Novel pH- and Temperature-Stable Pectate Lyase from Bacillus amyloliquefaciens S6 for Industrial Application.

    abstract::In this paper, we report cloning of a pectate lyase gene from Bacillus amyloliquefaciens S6 (pelS6), and biochemical characterization of the recombinant pectate lyase. PelS6 was found to be identical with B. subtilis 168 pel enzyme with 100% amino acid sequence homology. Although these two are genetically very close, ...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-019-00194-2

    authors: Bekli S,Aktas B,Gencer D,Aslim B

    更新日期:2019-09-01 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

  • Development of a new method for genetic transformation of the green alga Chlorella ellipsoidea.

    abstract::Chlorella ellipsoidea is a single-celled eukaryotic green microalgae with high nutritional value. Its value may be further increased if a simple, reliable and cost-effective transformation method for C. ellipsoidea can be developed. In this paper, we describe a novel transformation method for C. ellipsoidea . This sys...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-012-9554-3

    authors: Liu L,Wang Y,Zhang Y,Chen X,Zhang P,Ma S

    更新日期:2013-06-01 00:00:00

  • Enhanced protein secretion from insect cells by co-expression of the chaperone calreticulin and translation initiation factor eIF4E.

    abstract::Host protein synthesis is shut down in the lytic baculovirus expression vector system (BEVS). This also affects host proteins involved in routing secretory proteins through the endoplasmic reticulum (ER)-Golgi system. It has been demonstrated that a secretory alkaline phosphatase-EGFP fusion protein (SEFP) can act as ...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-012-9545-4

    authors: Teng CY,Chang SL,van Oers MM,Wu TY

    更新日期:2013-05-01 00:00:00

  • Clones identification and genetic characterization of Garnacha grapevine by means of different PCR-derived marker systems.

    abstract::This study uses PCR-derived marker systems to investigate the extent and distribution of genetic variability of 53 Garnacha accessions coming from Italy, France and Spain. The samples studied include 28 Italian accessions (named Tocai rosso in Vicenza area; Alicante in Sicily and Elba island; Gamay perugino in Perugia...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-010-9365-3

    authors: Meneghetti S,Costacurta A,Frare E,Da Rold G,Migliaro D,Morreale G,Crespan M,Sotés V,Calò A

    更新日期:2011-07-01 00:00:00

  • An improved protocol for the isolation of RNA from roots of tea (Camellia sinensis (L.) O. Kuntze).

    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

    authors: Muoki RC,Paul A,Kumari A,Singh K,Kumar S

    更新日期:2012-09-01 00:00:00

  • Expression of a Tardigrade Dsup Gene Enhances Genome Protection in Plants.

    abstract::DNA damage is one of the most impactful events in living organisms, leading to DNA sequence changes (mutation) and disruption of biological processes. A study has identified a protein called Damage Suppressor Protein (Dsup) in the tardigrade Ramazzotius varieornatus that has shown to reduce the effects of radiation da...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-020-00273-9

    authors: Kirke J,Jin XL,Zhang XH

    更新日期:2020-12-01 00:00:00

  • Expression and purification of human keratinocyte growth factor 2 by fusion with SUMO.

    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

    authors: Wu X,Nie C,Huang Z,Nie Y,Yan Q,Xiao Y,Su Z,Huang Y,Xiao J,Zeng Y,Tan Y,Feng W,Li X

    更新日期:2009-05-01 00:00:00

  • Miscanthus: genetic diversity and genotype identification using ISSR and RAPD markers.

    abstract::Due to the limited number of molecular studies focused on European gene pool investigation, it is necessary to perform plant material recognition. Eighteen accessions of three Miscanthus species, namely, M. × giganteus, M. sinensis, M. sacchariflorus were evaluated with the use of molecular marker systems such as: int...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-014-9770-0

    authors: Cichorz S,Gośka M,Litwiniec A

    更新日期:2014-10-01 00:00:00

  • Expression and purification strategies for the production of single-chain antibody and T-cell receptor fragments in E. coli.

    abstract::This work describes protocols for the production of single-chain antibody and T-cell receptor fragments in E. coli. A choice of methods is given for the purification of the recombinant fragments that rely on the use of either immunoaffinity or metal chelate affinity chromatography. The TCR fragments may have to be den...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/BF02779017

    authors: Molloy PE,Graham BM,Cupit PM,Grant SD,Porter AJ,Cunningham C

    更新日期:1995-12-01 00:00:00

  • Transfection efficiency along the regenerating soleus muscle of the rat.

    abstract::We investigated the efficiency of a single plasmid transfection along the longitudinal axis of the regenerating soleus of young rats. This also reflected transfection efficiency along the fibers because the soleus is a nearly fusiform muscle in young animals. The complete regeneration was induced by notexin and the tr...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-012-9555-2

    authors: Kósa M,Zádor E

    更新日期:2013-06-01 00:00:00

  • Rescue and production of vaccine and therapeutic adenovirus vectors expressing inhibitory transgenes.

    abstract::Expression of certain transgenes from an adenovirus vector can be deleterious to its own replication. This can result in the inhibition of virus rescue, reduced viral yields, or, in the worst case, make it impossible to construct a vector expressing the inhibiting transgene product. A gene regulation system based on t...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/BF02686012

    authors: Gall JG,Lizonova A,EttyReddy D,McVey D,Zuber M,Kovesdi I,Aughtman B,King CR,Brough DE

    更新日期:2007-03-01 00:00:00

  • Online resources for SNP analysis: a review and route map.

    abstract::The major online single nucleotide polymorphism (SNP) databases freely available as research tools for genetic analysis are explained, reviewed, and compared. An outline is given of the search strategies that can be used with the most extensive current SNP databases: National Centre for Biotechnology Information (NCBI...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章,评审

    doi:10.1385/mb:35:1:65

    authors: Phillips C

    更新日期:2007-01-01 00:00:00

  • Development of defined microbial population standards using fluorescence activated cell sorting for the absolute quantification of S. aureus using real-time PCR.

    abstract::In this article, four types of standards were assessed in a SYBR Green-based real-time PCR procedure for the quantification of Staphylococcus aureus (S. aureus) in DNA samples. The standards were purified S. aureus genomic DNA (type A), circular plasmid DNA containing a thermonuclease (nuc) gene fragment (type B), DNA...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-011-9417-3

    authors: Martinon A,Cronin UP,Wilkinson MG

    更新日期:2012-01-01 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

  • Studies on mRNA electroporation of immature and mature dendritic cells: effects on their immunogenic potential.

    abstract::Previous studies have shown that mRNA-electroporated dendritic cells (DCs) are able to process and present tumor-associated antigens, leading to the activation of tumor-specific T cells in vitro and in vivo. However, the optimal maturation state of antigen loading and half-life of the mRNA-translated protein product a...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-008-9071-6

    authors: Met O,Eriksen J,Svane IM

    更新日期:2008-10-01 00:00:00

  • Molecular Cloning, Expression and Characterization of Oenococcus oeni Priming Glycosyltransferases.

    abstract::Oenococcus oeni is the main bacterial species that drives malolactic fermentation in wine. Most O. oeni strains produce capsular exopolysaccharides (EPS) that may contribute to protect them in the wine hostile environment. In O. oeni genome sequences, several genes are predicted to encode priming glycosyltransferases ...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-017-0021-z

    authors: Dimopoulou M,Claisse O,Dutilh L,Miot-Sertier C,Ballestra P,Lucas PM,Dols-Lafargue M

    更新日期:2017-08-01 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

  • Reporter gene vectors and assays.

    abstract::Gene reporter systems play a key role in gene expression and regulation studies. This review describes the ideal reporter systems, including reporter expression vector design. It summarizes the many uses of genetic reporters and outlines the currently available and commonly used reporter systems. Each system is descri...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章,评审

    doi:10.1385/MB:13:1:29

    authors: Schenborn E,Groskreutz D

    更新日期:1999-11-01 00:00:00

  • Inter- and intra-varietal genetic variability in Malvasia cultivars.

    abstract::The DNA molecular analyses together with ampelography, ampelometry, and biochemistry are essential for grapevine identification and investigation of genetic differences among the Vitis vinifera L. cultivars and clones. Ten Malvasia cultivars (i.e., Istrian Malvasia; M. delle Lipari; M. bianca di Candia; M. di Candia A...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-011-9423-5

    authors: Meneghetti S,Poljuha D,Frare E,Costacurta A,Morreale G,Bavaresco L,Calò A

    更新日期:2012-03-01 00:00:00

  • New insights into protein-DNA interactions obtained by electron microscopy.

    abstract::The electron microscopic study of DNA-protein complexes can yield valuable information that is often not easily available by other methods. In this article we give a number of examples that were chosen to illustrate the utility of electron microscopy. Along with the strategy used are protocols that allow such experime...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1385/MB:16:1:77

    authors: Schnos M,Inman RB

    更新日期:2000-09-01 00:00:00

  • Cloning and nucleotide sequence of the thermostable beta-galactosidase gene from Pyrococcus woesei in Escherichia coli and some properties of the isolated enzyme.

    abstract::Pyrococcus woesei (DSM 3773) beta-galactosidase gene amplified by polymerase chain reaction was cloned into KpnI and HindIII binding sites of pET-30LIC expression plasmid. The obtained pGal2 (6785 bp) transcription vector was then transferred to Escherichia coli B121 (DE3) cells. High identity (99.9%) of DNA sequences...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/BF02740841

    authors: Dabrowski S,Maciuńska J,Synowiecki J

    更新日期:1998-12-01 00:00:00

  • Transgene methylation in mice reflects copy number but not expression level.

    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

    authors: Pena RN,Webster J,Kwan S,Korbel J,Whitelaw BA

    更新日期:2004-03-01 00:00:00

  • Biological significance of RNA editing in cells.

    abstract::RNA editing is one of the post-transcriptional RNA processes. RNA editing generates RNA and protein diversity in eukaryotes and results in specific amino acid substitutions, deletions, and changes in gene expression levels. Adenosine-to-inosine RNA editing represents the most important class of editing in human and af...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章,评审

    doi:10.1007/s12033-012-9498-7

    authors: Tang W,Fei Y,Page M

    更新日期:2012-09-01 00:00:00

  • GM risk assessment.

    abstract::GM risk assessments (GMRAs) play an important role in the decision-making process surrounding the regulation, notification and permission to handle Genetically Modified Organisms (GMOs). Ultimately the role of each GMRA will be able to ensure the safe handling and containment of the GMO; and to asses any potential imp...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章,评审

    doi:10.1007/s12033-009-9237-x

    authors: Sparrow PA

    更新日期:2010-03-01 00:00:00

  • Stable Expression of Adalimumab in Nicotiana tabacum.

    abstract::Production of monoclonal antibodies and pharmaceutical proteins in transgenic plants has been the focus of many research efforts for close to 30 years. Use of plants as bioreactors reduces large-scale production costs and minimizes risk for human pathogens contamination. Stable nuclear transformation of the plant geno...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-018-0075-6

    authors: Zvirin T,Magrisso L,Yaari A,Shoseyov O

    更新日期:2018-06-01 00:00:00

  • Molecular chaperone-assisted production of human alpha-1,4-N-acetylglucosaminyltransferase in silkworm larvae using recombinant BmNPV bacmids.

    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

    authors: Nakajima M,Kato T,Kanamasa S,Park EY

    更新日期:2009-09-01 00:00:00

  • Surface display of a bifunctional glutathione synthetase on Saccharomyces cerevisiae for converting chicken feather hydrolysate into glutathione.

    abstract::The low economic profits of feather recycling lead that the large amount of feathers is currently discarded in China. To convert feather hydrolysates into GSH with high values, surface display of the bifunctional glutathione synthetase encoded by gcsgs from Streptococcus thermophilus on Saccharomyces cerevisiae and th...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-014-9750-4

    authors: Qiu Z,Tan H,Zhou S,Cao L

    更新日期:2014-08-01 00:00:00

  • The Interacting Domains of PREP1 and p160 are Endowed with a Remarkable Structural Stability.

    abstract::PREP1/p160 is a protein complex with relevant physiopathological roles in vivo. p160 regulates PREP1 transcriptional activity by preventing the formation of other PREP1-containing complexes, whereas PREP1 regulates p160 activity by increasing its stability. This induces the repression of the insulin-regulated glucose ...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-016-9932-3

    authors: Lorenzo V,Mascanzoni F,Vitagliano L,Ruvo M,Doti N

    更新日期:2016-05-01 00:00:00