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Construction and Analysis of a Subtractive cDNA Library on Early Embryonic Development Related Genes in Ducks

Author: ZhongLingXiu
Tutor: LuLiZhi;ShiFangXiong
School: Nanjing Agricultural College
Course: Animal Genetic Breeding and Reproduction
Keywords: Duck Early embryonic development DSN Subtractive hybridization cDNA library
CLC: S834
Type: Master's thesis
Year: 2010
Downloads: 1
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Abstract


In this experiment, we use the Jinyun duck embryo as the experimental material, and use a DSN mediated normalization subtractive hybridization method to construct a subtracted cDNA library of duck early embryonic development-related genes. We hope to find the related new genes, exploring the molecular mechanism of embryonic development-related genes expression, and establish the pattern of gene expression in different developmental stages.In this study, two subtractive hybridization librarys that related early embryonic development of ducks were constructed. In the first library, two different periods (6 hours and 20 hours) of duck embryo were used as the driver and the tester respectively. A DSN-mediated subtractive hybridization method was employed, then we connect the reduction products with a pUCm-T carrier, through the blue-white screening,216 positive clones were randomly selected, and finally selected fragments over 750bp by bacilli PCR and then sequenced. Removing the vector sequence on Vecscreen and the redundant sequence with Staden Package. Finally we obtained a total of 54 independent differentially expressed genes, analysis with BLAST,44 genes of which have homologous genes, another 10 with no homologous genes. Through the GO annotation, we found these genes play an important role in various biological pathways, such as the metabolism, the establishment of cellular localization, biological regulation, and some genes may participate in two or more biological pathways. Through further research of the related homologous genes, we found some genes play an important role in neural development, immune regulation, transcription regulation, and so on, while the functions of other genes have not been reported yet. In the second library,6 hours and 20 hours duck embryo were used respectively, as the driver and the tester, using the same method as above,66 differentially expressed genes were obtained,48 of which have homologous genes, another 18 has no homologous genes.27 of these 66 genes are the same with the genes screened in the first gene library.23 of which have homologous genes and 4 have no homologous genes, indicating that these genes continued functioning during early embryonic development. And there are still 25 genes may be specific expression at this stage, some of which genes are still related with neural development and immune regulation.Some new genes are found in two libraries, these new genes and the genes which the functions has not been explored, could available through RNA interference technology in further studies, to make a better understanding about the process of the poultry early embryonic development.

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