| In plant cells, except nucleus, chloroplast and mitochondria also contain genetic material DNA. With the development DNA sequencing technology, the number of the plants species whose nuclear, chloroplast and mitochondrial genome sequencing projects have been completed is steadly increasing recently. Studie on the genome structure and gene information in nucleus, chloroplasts and mitochondria by the combination method of bioinformatics and biological experiments could provide important reference on the genetic relationship among different species, the origin of plant species and their evolution. Artificial microRNAs (amiRNA) gene silencing technology is considered as a efficient method for studying the mechanism of coordination gene expression between nuclear and organellar genomes.Cytochrome b (cob) is one of the important component in respiratory chain, which is encoded both by nuclear and mitochondrial genome each with only one copy in Arabidopsis thaliana. Cob gene knock down by artificial microRNAs gene silencing technology should help to reveal the mechanism of coordinated regulation of cob gene expression between nuclear and organellar genomes. In the present research, cob silencing vectors for nucleus genome were constructed by amiRNA technology , and were transformed into A.thaliana via Agrobacterium-mediated method.The main results are as follows:1) Through analyzing the features of the natural miRNA and the candidates provided by WMD3, amicroRNA of the nucleus cob gene was designed.2) Nucleus cob silencing vectors was constructed using the templates synthesed by overlapping PCR method.3) Nuclear cob silencing vectors were transformed into A.thaliana via Agrobacterium-mediated method.In conclusion, A.thalianan cob silencing vectors was successfully constructed. The results of present research has paved the way for studying the mechanism of coordinated regulation of cob gene expression between nuclear and organellar genomes. |
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