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Chicken small RNA library construction and the size of the non-coding RNA screening

Author: HuangShouJun
Tutor: ZhuDaHai
School: Peking Union Medical College , China
Course: Biochemistry and Molecular Biology
Keywords: Non-coding RNA Chicken Expression profiling MuN121
CLC: S831
Type: Master's thesis
Year: 2008
Downloads: 217
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Abstract


Non - coding RNA plays a very important role in the activities of life , the processing involved in the regulation of chromatin structure , transcription and transcription regulation , protein synthesis , transport and the activity of regulation , but the non- coding RNA known only account biological body all non- coding RNA fraction , the vast majority of non- coding RNA has yet to be found , and functional studies . By constructing a special cDNA library is found that the new non-coding RNA is a more effective method . In this study, the chicken as a model organism , build a library of non- coding RNA enriched small molecules to find a large number of new non-coding RNA and Northern hybridization experiments to verify the presence of these non-coding RNA . The integrity of these new non-coding RNA research gives us a deeper understanding of non-coding RNA . First , through the analysis of a variety of non-coding RNA expression in the different tissues of the chicken and skeletal muscle in different developmental stages , it is found that the vast majority of non-coding RNA has a different level of expression in time and space , these non-coding RNA in vivo generation by strict regulation of degradation , suggesting that they have an important role in certain life activities ; chicken in many non- coding RNA in mice and humans and there is no presence of homologous RNA molecules genome level than on can not find homologous sequences, which will generate a considerable number of non-coding RNA species-specific organism in evolution , with the protein-coding genes have conserved compared to having a significantly different , these non-coding RNA may very large extent determine the uniqueness of the species . This experiment also found a chicken skeletal and cardiac muscle expression of non-coding RNA - MuN121 Blast analysis shows that it is being located in the intron of a known protein-coding genes , Northern hybridization showed MuN121 and the host gene expression at the RNA level have consistent spectral . In order to study the generation of MuN121 independent transcription from or from pre-mRNA processing comes , we constructed two kinds of wild-type and the intron 5 ' end of the splice donor site mutation carrier transfected 293T cells found mutant not produce ncRNA wild type can produce . Show that generation of MuN121 depends on the success of pre-mRNA splicing MuN121 and host genes regulated by the same transcription mechanism .

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Poultry > Chicken
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