Background:In the recent decade, the reverse genetics method has been broadly used for rescue of negative-stranded RNA viruses from cDNA or viral minigenomes. This technique has been applied to study different steps in virus replication and virus-host interactions. Reverse genetics could also be implemented for design of new vaccines. The T7 RNA polymerase activity as well as virus (nucleocapsi...

78 Abstract Pneumoviruses are responsible for significant respiratory disease in their hosts and represent a major problem for human and animal health. Pneumoviruses are members of the family Paramyxoviridae, subfamily Pneumovirinae and the virus particles consist of a negative-sense, nonsegmented RNA genome within a helical nucleocapsid structure enveloped in a lipid membrane derived from the ...

Reverse genetics consists in the modification of the activity of a target gene to analyse the phenotypic consequences. Four main approaches are used towards this goal and will be explained in this review. Two of them are centred on genome alterations. Mutations produced by random chemical or insertional mutagenesis can be screened to recover only mutants in a specific gene of interest. Alternat...

The genetics of viruses are determined by mutations of their nucleic acid. Mutations can occur spontaneously or be produced by physical or chemical means: for example, the application of different temperatures or mutagens (such as hydroxylamine, nitrous acid, or alkylating agents) that alter the nucleic acid. The classic way to study virus mutants is to identify a change in phenotype compared w...

The number of sequenced genes whose function remains unknown continues to climb with the continuing decrease in the cost of genome sequencing. Comparative genetics and bioinformatics have been invaluable in investigating the function of the genes that have been sequenced, but the elucidation of gene function in planta remains a huge challenge. Many gene functions have been defined through the u...

The zebrafish has become a popular model system for the study of vertebrate developmental biology because of its numerous strengths as a molecular genetic and embryological system. To determine the requirement for specific genes during embryogenesis, it is necessary to generate organisms carrying loss-of-function mutations. This can be accomplished in zebrafish through a reverse genetic approac...