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Construction of Silence Vector for Cytochrome b Gene in Nucleus of Arabidopsis Thaliana and the Transformation

Author: ZhouZuo
Tutor: HeGuangYuan；YangGuangXiao
School: Huazhong University of Science and Technology
Course: Biomedical Engineering
Keywords: Arabidopsis Cytochrome b gene RNA interference Synthetic miRNA
CLC: Q943
Type: Master's thesis
Year: 2011
Downloads: 12
Quote: 0
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Abstract


Plant cells, in addition to the nucleus , chloroplasts and mitochondria also contain genetic material DNA . With the genome sequencing technology continues to improve , the plant has completed the nuclear genome , increasing the number of species in the chloroplast genome and mitochondrial genome sequencing . By bioinformatics analysis method and biological experiments phase combination of methods to study the chloroplast genome , the mitochondrial genome and the nucleus gene group three of the structure and sequence information characteristics can reveal gene transfer and the origin of species , the law of evolution ; addition also for the different species phylogenetic relationships among other aspects of the study provide important theoretical basis . The synthetic miRNA interference technology relationship between the regulation of gene expression provides effective research methods for the study of the nucleus and organelles . Cytochrome b (cytochrome b, cob) is one of the important members of the respiratory chain . Cytochrome b gene in Arabidopsis thaliana nuclear genome and the mitochondrial genome exist , and are single-copy genes using RNA interference technology to cut the cob in the nuclear genome gene expression can clear nucleus and mitochondria of the gene expression regulation relationship. The method of the present study, synthetic miRNA construct cob gene in the genome of Arabidopsis nucleus interference carrier , and the Arabidopsis transformation . To obtain the main results are as follows : 1 ) by analyzing the parameters of the endogenous miRNA select target genes , the candidate sequences reference WMD3 provide complete cytochrome the b gene amiRNA molecular design . 2) According to the completion of the precursor of the selected primer design. Successfully synthesized by overlap extension required template and product ; to use overlap extension PCR synthesized amiRNA precursor . The sequencing results show that , the synthesis amiRNA fully comply with the design requirements . 3) interference carrier into Arabidopsis by Agrobacterium anther dip method . The results of this study to explore the nucleus and mitochondria cob gene expression and regulation of the relationship and differences laid the initial foundation .

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CLC: > Biological Sciences > Botany > Plant Cell Genetics
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