Loss of Function by Mutation Approach, Genomic Sequences, Genomic Scale, Insertion Mutagenesis, Agrobacterium Plasmid, DNA Sequences

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“Loss of function by mutation” approach.

Since search in the databases does not resolve functions of many genomic sequences, mutations for loss of function are artificially produced in large number at the genomic scale. Thee mutations are then used for isolation of genes followed by the study of functions of these genomic sequences. This leads to discovery of new gene and their functions. Several approaches are available for producing these mutations for loss of function.

Insertion mutagenesis.

Insertion mutagenesis is a very powerful approach for identification and isolation of gene and for the identification of their function. It involved insertion of transposons. Or T-DNA from Agrobacterium plasmid on random sites followed by the study of ‘loss of function’ mutants thus obtained. The DNA sequences with inserted transposon or T-DNA are then isolated inserted transposon or T-DNA are then isolated and the structure of the DNA is studied and related with the function that was disrupted in the mutant. A number of such studies have been conducted.

Insertional Mutagenesis for Annotation of Genomic Sequences through 'Loss of Function'

Insertional Mutagenesis for annotation of Genomic Sequences through 'Loss fo function'

Insertional Mutagenesis for annotation of Genomic Sequences through 'Loss of function2'

(b) Targeted gene disruption.

Targeted gene disruption discussed earlier in 7 has also been used for study of gene function.

(c) Gene silencing.

Gene silencing can also be achieved using antisense or sense suppression of gene function or using dsRNA for interference (RNAi). This has also been used for study of gene function.

Gene trap and enhancer trap.

Gene trap and enhancer trap are two approaches, in which a gene construct carrying a marker gene is inserted. If the marker gene is expressed this is taken to mean that a promoter/enhancer sequence or a gene with a promoter sequence is present at the site of insertion.

Gene Trap and Enhancer Trap approaches for annotation of Genome Sequences through expression of a reporter gene

Gene Trap and Enhancer Trap approaches for annotation of genome sequences through expression of a reporter gene

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