Welcome To Website IAS

Hot news
Achievement

Independence Award

- First Rank - Second Rank - Third Rank

Labour Award

- First Rank - Second Rank -Third Rank

National Award

 - Study on food stuff for animal(2005)

 - Study on rice breeding for export and domestic consumption(2005)

VIFOTEC Award

- Hybrid Maize by Single Cross V2002 (2003)

- Tomato Grafting to Manage Ralstonia Disease(2005)

- Cassava variety KM140(2010)

Centres
Website links
Vietnamese calendar
Library
Visitors summary
 Curently online :  13
 Total visitors :  7683924

Positive-negative-selection-mediated gene targeting in rice.
Monday, 2015/01/26 | 08:04:22

Shimatani Z, Nishizawa-Yokoi A, Endo M, Toki S, Terada R.

 

Front Plant Sci. 2015 Jan 5;5:748. doi: 10.3389/fpls.2014.00748. eCollection 2014.

http://www.ncbi.nlm.nih.gov/pubmed/25601872

 

Abstract

 

Gene targeting (GT) refers to the designed modification of genomic sequence(s) through homologous recombination (HR). GT is a powerful tool both for the study of gene function and for molecular breeding. However, in transformation of higher plants, non-homologous end joining (NHEJ) occurs overwhelmingly in somatic cells, masking HR-mediated GT. Positive-negative selection (PNS) is an approach for finding HR-mediated GT events because it can eliminate NHEJ effectively by expression of a negative-selection marker gene. In rice-a major crop worldwide-reproducible PNS-mediated GT of endogenous genes has now been successfully achieved. The procedure is based on strong PNS using diphtheria toxin A-fragment as a negative marker, and has succeeded in the directed modification of several endogenous rice genes in various ways. In addition to gene knock-outs and knock-ins, a nucleotide substitution in a target gene was also achieved recently. This review presents a summary of the development of the rice PNS system, highlighting its advantages. Different types of gene modification and gene editing aimed at developing new plant breeding technology based on PNS are discussed.

 

FIGURE 1. Schematic diagram of various gene modifications by PNS-mediated GT. (A) The brown box indicates the gene to be targeted on a genome sequence shown as black lines. The brown arrow represents the promoter of the gene. (B) PNS vector for GT. The green arrows are the negative markers; the red arrow is the positive marker. The pink box is the transcriptional stop sequence of En/Spm. The gray arrows are loxP sequences. Double-headed arrows under the vector indicate the homology regions for HR. The blue line is T-DNA sequence. (C) HR process for GT between the target gene and PNS vector. The thick lines of black and blue indicate newly synthesized DNA sequences in genome and T-DNA, respectively. (D) Gene knock-out of the target gene by insertion of a positive marker with En/Spm, which can be removed via subsequent Cre-loxP recombination caused by introduced Cre gene (yellow arrow). (E) Reactivation of knock-out gene in (D) by Cre-loxP recombination. (F) Nucleotide(s) substitution (red lines), insertion, and deletion in the target gene can be induced by designing a homology arm in the PNS vector in (B) and subsequent positive marker elimination by Cre-loxP recombination in (D). (G) Gene knock-in modification where the endogenous promoter sequence is connected to the GUS coding sequence (indicated as a blue box with En/Spm). (H) Gene knock-in modification where the mOrange coding sequence, indicated as an orange box, is connected precisely to the stop codon of the target gene; both endogenous promoter activity and protein localization of the target gene are detectable. (I–K) Diagrams of segregated plants from knock-in T0 into homozygote (I), heterozygote (J), and wild type (K). GUS expression image as blue leaves is shown in (I,J). Dwarf phenotype in (I) is a reflection of the disrupted target gene.

Back      Print      View: 1089

[ Other News ]___________________________________________________
  • Genome-wide analysis of autophagy-associated genes in foxtail millet (Setaria italica L.) and characterization of the function of SiATG8a in conferring tolerance to nitrogen starvation in rice.
  • Arabidopsis small nucleolar RNA monitors the efficient pre-rRNA processing during ribosome biogenesis
  • XA21-specific induction of stress-related genes following Xanthomonas infection of detached rice leaves.
  • Reducing the Use of Pesticides with Site-Specific Application: The Chemical Control of Rhizoctonia solani as a Case of Study for the Management of Soil-Borne Diseases
  • OsJRL, a rice jacalin-related mannose-binding lectin gene, enhances Escherichia coli viability under high-salinity stress and improves salinity tolerance of rice.
  • Production of lipopeptide biosurfactants by Bacillus atrophaeus 5-2a and their potential use in microbial enhanced oil recovery.
  • GhABF2, a bZIP transcription factor, confers drought and salinity tolerance in cotton (Gossypium hirsutum L.).
  • Resilience of cassava (Manihot esculenta Crantz) to salinity: implications for food security in low-lying regions.
  • Cellulose synthase complexes act in a concerted fashion to synthesize highly aggregated cellulose in secondary cell walls of plants
  • No adverse effects of transgenic maize on population dynamics of endophytic Bacillus subtilis strain B916-gfp
  • Identification and expression analysis of OsLPR family revealed the potential roles of OsLPR3 and 5 in maintaining phosphate homeostasis in rice
  • Functional analysis of molecular interactions in synthetic auxin response circuits
  • Titanium dioxide nanoparticles strongly impact soil microbial function by affecting archaeal nitrifiers.
  • Inducible Expression of the De-Novo Designed Antimicrobial Peptide SP1-1 in Tomato Confers Resistance to Xanthomonas campestris pv. vesicatoria.
  • Toward combined delignification and saccharification of wheat straw by a laccase-containing designer cellulosome
  • SNP-based discovery of salinity-tolerant QTLs in a bi-parental population of rice (Oryza sativa)
  • Pinpointing genes underlying the quantitative trait loci for root-knot nematode resistance in palaeopolyploid soybean by whole genome resequencing.
  • Transcriptome- Assisted Label-Free Quantitative Proteomics Analysis Reveals Novel Insights into Piper nigrum -Phytophthora capsici Phytopathosystem.
  • Brassinosteroids participate in the control of basal and acquired freezing tolerance of plants
  • Rapid hyperosmotic-induced Ca2+ responses in Arabidopsis thaliana exhibit sensory potentiation and involvement of plastidial KEA transporters

 

Designed & Powered by WEBSO CO.,LTD