Map Based Cloning for Gene Isolation,Mutation,Chromosome,Transformation of Mutant,Cloning Utilizing,Cystic Fibrosis

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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

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Map Based Cloning for Gene Isolation
Map based cloning can be used for isolation of a gene for which a mutation can be identified and in a crop for which a saturated molecular map is available. Availability of comprehensive YAC or BAC. Genomic library is another requirement. Most of these requirements are met in major crop plants. The mutation is first mapped close to any of the molecular markers that are already mapped. A variety of molecular markers are available for the preparation of these molecular maps (e.g., RFLP, RAPD, SSR, STMS, AFLP, SNP, etc.; consult next chapter for molecular markers).

Different Steps involved in isolation of gene through map based cloning based on a RFLP based molecular map

Different Steps Involved in Isolation of Gene Through Map Based Cloning Based on a RFLP Based Molecular Map

Once the mutant is mapped, the most closely linked pair of flanking markers are used as hybridization probes to isolate clones containing the genomic segment located between the markers. Techniques of chromosome walking, chromosome jumping and chromosome landing can be used to reach closer to the gene of interest and to reach closer to the gene of interest and to isolated. These probable clones can be used for transformation of the mutant, to find out which of the clones complements the mutation. This clone then should contain the gene. Using this approach, several genes have been isolated in crop plants and animals including humans.A gene for cystic fibrosis in humans was cloned through map based cloning utilizing the technique will be increasingly used in future fo isolation of a variety of genes for which the gene products are not known.

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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