A Study On Alternative Splicing Variants Of The Genes Associated With Hypoxia In C. Elegans

Oxygen is the basis of the activities of organisms to sustain life, lacking oxygen will cause injury or hypoxia-related diseases in the body. Organisms adapt hypoxic stress by regulating the expression of hypoxia-related genes. Alternative splicing has emerged as a key mechanism that accounts for gene expression diversity in metazoan organisms implicated in many diseases including cancer. Many hypoxia-related genes involve in hypoxia response in different splice variants by alternative splicing and play an important role. Increasing evidence suggests that low-abundant gene may play fundamental roles in biological processes. However, we known a little about the low-abundant gene because of their small mass and high heterogeneity. To investigate the new alternative splicing of different abundance genes, and then identify the relationship between the alternative splicing and human healthy, we do the following researches:Firstly, by using the model organism C. elegans as the material, and using reverse transcription by GSPs and nested PCR technique, we identify the splicing variants of genes (such as HIF-1,egl-9,glb-3,glb-5,glb-8,glb-13,glb-18, and etc. ) and discoverede a new splicing variants of glb-18. By comparing with normal techniques, this method can detect splicing variants efficiently, and provide more new reference data for the transcriptome and alternative splicing.Secondly, using reverse transcription by GSPs and nested PCR techniques, we detect the variety of genes . The results showed that the expression of all genes we tested were changed. The result suggests that there is a closer relationship between alternative and hypoxia, furthermore, it also certified that genes play different roles in different developmental stage and various environment.Thirdly, because of the low transcript and the high heterogeneity, we make different abundance gene(actin-1,egl-9,hif-1,glb-18 and TrkC) of model organism C. elegans and mouse as the material, using two steps of reverse transcription, to research the efficiency of the gene's expression. The result shows that this method could not only increase the mass of the low transcripts, but also provided a laid an important foundation for further study of alternative splicing of low abundance gene.In summary, the study has provided new reference methods for the research of alternative splicing and low abundance gene, and also provided important clues to study the gene which is closely connected with disease hypoxia-related.