Promoter, Enhancer and Gene Trap for Isolation of Genes,Transformants,T-DNA,Clone,Tobacco,Mutation,Phenotypic Change

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Isolation, Sequencing and Synthesis of Genes Isolation, Sequencing and Synthesis of Genes - Introduction Isolation of Genes Isolation of Genes Coding for Specific Proteins Isolation of Genes with Known or Unknown Products having Tissue Specific Expression Isolation of Genes with Known or Unknown Products Using DNA or RNA Probes Use of Heterologus Probes Use of Synthetic Probes Isolation of Genes Coding Use of Transposable Elements Transposon Tagging T-DNA-Insertion Mutagenesis for Isolation of Plant Genes Promoter Enhancer and Gene Trap for Isolation of Genes Differential Screeing and Differential Display Technique for Isolation of Genes Subtractive Hybridizatioon for Gene Isolation Map Based Cloning for Gene Isolation Isolation of Novel Genes by Asis Data at JNU New Delhi

Home >> Biotechnology and Genomics >> Isolation, Sequencing and Synthesis of Genes >> Promoter, Enhancer and Gene Trap for Isolation of Genes

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Promoter, Enhancer and Gene Trap for Isolation of Genes
Although transformants for a large number of traits have already become available, all genes are not tagged in these transformants. Therefore, a major effort is needed to isolate any new gene that is not tagged in any of the available transformants. A large number of transformed plants will have to be produced and propagated to isolate these new genes. Furthermore, the insertion of transposon or T-DNA may nor result in perceptible phenotypic change. Therefore, if a new gene (for which mutation could not be produced so far through transposon tagging or T-DNA insertion) needs to be isolated, one may like to use a new strategy to clone the gene of interest.

One such strategy involves the insertion of a reporter gene with or without a TATA box or a splice acceptor (SA). In using a hypothetical gene three such strategies are illustrated, which are described as ‘enhancer trap’. ‘promoter trap’ and ‘gene trap’. In each case, the reporter gene will be expressed under the influence of enhancer/promoter sequence of a gene of unknown function and can be used as a tag for the isolation of the gene involved. In tobacco, this technique has been successfully used for identification of a gene involved in auxin synthesis by growing cultures on a medium not containing auxin.

Promoter enhancer and gene trap stragies for identification and isolation of genes with unknown functions

Mutant complementation
In this technique, DNA close from the wild type strain are selected which should be able to complement the mutants, which are thus transformed into wild type. Once these DNA clones are available, the protoplasts derived from the mutant plant may be transformed using wild type clones and the transgenic plants are produced. Once this is done, the gene of interest can be isolated from DNA extracted from the transformed plants using wild type complementary clone as a probe.

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Sequencing of a Gene or a DNA Fragment Maxam and Gilbert's Chemical Degradation Method Sanger's Dideoxynucleotide Synthetic Method Automatic DNA Sequencers Slab Gel Versus Capillary Sequencers Slab Gel Sequencing Systems Capillary Gel Sequencing Systems Direct DNA Sequencing Using PCR Ligation Mediated PCR LMPCR DNA Sequencing Through Transcription DNA Sequencing by Hybridization Using Microarrays on DNA Chips DNA Sequencing by DE-MALDI-TOF Mass Spectrometry Synthesis of Genes Synthesis of Gene for Yeast Alanyl tRNA Synthesis of Oligonucleotides Synthesis of Three Duplex Fragments Synthesis of Gene from Three Duplex Fragments Synthesis of Gene for a True Precursor tRNA Total Synthesis of a Human Leukocyte Interferon Gene Gene Synthesis Machines Protection Chemistry and Amidites Permanent Protecting Groups Temporary Protecting Groups Synthesis of Gene Using PCR Synthesis of Genes From mRNA