Caenorhabditis Elegans Lab Report
Abstract The body of Caenorhabditis elegans (C. elegans) are surrounded by a cuticle that is molted four times in development although not all cuticle collagen genes are expressed at once. Mutations in these genes lead to morphological defects such as blister formation in the cuticle. The blister phenotype (Bli) is inherited recessively and develop in the adult stage of C. elegans. Other cuticle collagen mutations, such as the uncoordinated (unc), roller (rol) and dumpy (dpy) mutations, can suppress the Bli phenotype. Sc109 is also another mutation that is found in a carrier of the bli-1 allele and was found to incompletely suppress the formation of blisters. Tetraspanin protein (TSP-15) is found to be needed to maintain the integrity of the epithelial membrane as a deficiency of TSP-15 lead to separation of cuticle layers. Dpy-5 alleles are dominant suppressors of bli-4 mutations although suppression is incomplete. Low counts of Bli phenotype emerged in a population of C. elegans along with multiple unc mutants indicating incomplete suppression of bli mutation. Disorders like epidermolysis
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The technique gives a high frequency of point mutations that is distributed randomly in the genome (Kurowska et al., 2011). The high throughput strategy uses a chemical mutagen, Ethyl methane sulfonate (EMS), and a technique that identifies single base changes within the target gene. With the TILLING method, multiple alleles are amplified by PCR to for DNA heteroduplexes which are double stranded nucleic acid. When it is heated and cooled, a bubble forms where two DNA strands are mismatched and is cleaved by single stranded nucleases. Mismatches can be because of induced mutation or natural variation (Henikoff, Till, & Comai, 2004). The cleaved products are run on a gel to be separated by
The technique gives a high frequency of point mutations that is distributed randomly in the genome (Kurowska et al., 2011). The high throughput strategy uses a chemical mutagen, Ethyl methane sulfonate (EMS), and a technique that identifies single base changes within the target gene. With the TILLING method, multiple alleles are amplified by PCR to for DNA heteroduplexes which are double stranded nucleic acid. When it is heated and cooled, a bubble forms where two DNA strands are mismatched and is cleaved by single stranded nucleases. Mismatches can be because of induced mutation or natural variation (Henikoff, Till, & Comai, 2004). The cleaved products are run on a gel to be separated by