In some occasion, especially when there is food deprivation or if c. Elegans is growing in a plate at the laboratory, it can take an alternative form of life known as the dauer larval stage as shown in Figure 1. Dauer larvae cannot eat since their mouths are plugged and also it has a thin body. Remarkably, dauers can stay alive for three months. However, they seem to be non-aging because they live for months until they consume food. It is after taking food that dauers enter the L4 stage where they live for more than the normal c.
Elegans. It is because of this that c. Elegans leads to further research regarding anti aging for human benefits. Promoter trapping Promoter trapping was developed before genome sequence project. It was used to screen the gene expression within an organism. In c. Elegans, a reporter bacterial gene called lacZ was involved with encoding of the enzyme ß-galactosidase. This enzyme is fused to the regulatory elements of different genes including c. Elegans that is expresses cells by ß-galactosidase (Finney M, 2012, pp. 1-2).
In order to generate a promoter trapping in c. Elegans, DNA fragment need to be extracted from c. Elegans by restriction enzyme. It is then transferred to a reporter gene that has or has no promoter. Following this mixture of DNA and reporter, it induces to host organism or in this case into c. Elegans to observe the expression (Kulkarni, 2008). In this practical, both UL1 and UL6 are already fused with lacZ gene in their regulatory elements. This is used to determine the site of expression by histochemical staining.
The histochemical staining is seen in this practice X-gal. This staining is colourless substrate of ß- galactosidase. When it is cleaved, it gives an insoluble blue product that is a precipitate at the sites of ß-galactosidase activity. Materials and methods This study was done in pairs such that two strains of the nematode worm Caenorhabditiselegans, UL1 and UL6 are provided for each pairs, each student worked with one stain (Kulkarni, 2008, p. 12). Both of the worms stain is washed separately by distilled water using eppendorf tube for 5 minutes.
Later the worms are settled at the bottom of tube where they each are transferred to a multiwell glass or microscope slide. Each worm slide is labelled and then covered with coverslip. UL1 and UL6 slides are then placed on a metal plate on dry ice to freeze the worm and penetrate the stain. 3 minutes later, the slides are transferred quickly to an acetone at -20 degrees Celsius.
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