Reserve genetics is a tool used to identify the functionality of a known gene sequence or a gene family by genetic editing to generate the mutant phenotype. As it is done in a direction from gene to phenotype, which is opposite to ‘forward genetics’, hence named as reverse genetics. As the whole genome sequences are much more accessible nowadays with various genome sequencing technologies developed. This enables modifying a known gene, and to create a known mutation. The mutated gene might then give a change in the phenotype. We can then link this phenotypic change back to the original gene to determine the functionality of the original gene. (1-7)
With reserve genetic approaches, there are three major ways of modifying a gene sequence, deletion, …show more content…
The evolution of technologies offered more variety of genes been analyzed, which helped reserve genetic approaches becoming more accessible and underlying genetic functions with expanding of organism models like animals and virus. (1-5) Moreover, the expanded studies with the animal and virus model give insights into the invention of vaccines and medical treatments. (6) Although reserve genetics approaches have become more and more mature in functional genomics, the improvement of these techniques is still needed. For instance, lowering the cost of each test by taking advantages of NGS technology, so that the reserve genetics can be used more widely across the industries with more genes were analyzed. (1,3)
T-DNA insertion technique is one of the insertional mutagenesis techniques that has been widely used in both forward and reverse genetic approaches (1,4) and it is one of the most widely used techniques in reverse genetic approaches. (2-3) This technique uses T-DNA as a foreign gene to transfer itself into target genome, thus to disrupt the function of the target genome. One of the advantages of using T-DNA insertion is that the T-DNA contains recognizable tags that can be detected later to identify the mutant is introduced by insertion. …show more content…
Polymerase chain reaction (PCR) is used to screen the insertions in the genomes. (2,4) After the insertions are confirmed, the PCR process will be repeated in conjunction with the uses of target gene-specific primer and T-DNA specific primers. This step is to help with identifying the insertions located within or close to the desired region. (2, 6) Once the inserts are identified, the targeted gene sequence needs to be isolated by flanking sequence (7) and the mutant needs to be confirmed. (2) However, without phenotypic analysis, the linkage between the produced phenotype and mutant is remaining unclear. Thus, it is essential to confirm that the changes in mutant phenotype are caused by the insertion. (2) Some of the methods available to make the confirmation is co-segregation with reference, or transgenic technology to reproduce an identical mutant from wide type.
With reserve genetic approaches, there are three major ways of modifying a gene sequence, deletion, …show more content…
The evolution of technologies offered more variety of genes been analyzed, which helped reserve genetic approaches becoming more accessible and underlying genetic functions with expanding of organism models like animals and virus. (1-5) Moreover, the expanded studies with the animal and virus model give insights into the invention of vaccines and medical treatments. (6) Although reserve genetics approaches have become more and more mature in functional genomics, the improvement of these techniques is still needed. For instance, lowering the cost of each test by taking advantages of NGS technology, so that the reserve genetics can be used more widely across the industries with more genes were analyzed. (1,3)
T-DNA insertion technique is one of the insertional mutagenesis techniques that has been widely used in both forward and reverse genetic approaches (1,4) and it is one of the most widely used techniques in reverse genetic approaches. (2-3) This technique uses T-DNA as a foreign gene to transfer itself into target genome, thus to disrupt the function of the target genome. One of the advantages of using T-DNA insertion is that the T-DNA contains recognizable tags that can be detected later to identify the mutant is introduced by insertion. …show more content…
Polymerase chain reaction (PCR) is used to screen the insertions in the genomes. (2,4) After the insertions are confirmed, the PCR process will be repeated in conjunction with the uses of target gene-specific primer and T-DNA specific primers. This step is to help with identifying the insertions located within or close to the desired region. (2, 6) Once the inserts are identified, the targeted gene sequence needs to be isolated by flanking sequence (7) and the mutant needs to be confirmed. (2) However, without phenotypic analysis, the linkage between the produced phenotype and mutant is remaining unclear. Thus, it is essential to confirm that the changes in mutant phenotype are caused by the insertion. (2) Some of the methods available to make the confirmation is co-segregation with reference, or transgenic technology to reproduce an identical mutant from wide type.