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標題: 實體定位與結構分析四個玉米B染色體專一性RAPD標誌
Physical mapping and structural analysis of four B-chromosome specific RAPD markers in maize
作者: 高國維
Kao, Kuo-Wei
關鍵字: 玉米;Maize;B染色體;RAPD;B-chromosome;RAPD
出版社: 農藝學系所
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B染色體 (B-chromosome) 又稱為超數染色體 (supernumerary chromosome),是指在個體中所帶有不屬於正常染色體 (A chromosomes) 之非必要的額外染色體。玉米B染色體為末端中節染色體,其長臂可以細分為近端異染色質 (proximal heterochromatin)、近端常染色質 (proximal euchromatin)、遠端異染色質 (distal heterochromatin) 及遠端常染色質 (distal euchromatin)。玉米B染色體的DNA組成與A染色體相似,因此造成玉米B染色體的研究困難,目前僅發現三個玉米B染色體專一序列 (ZmBs、CL-repeat及StarkB)。本研究對四個B染色體專一性RAPD (random amplification of polymorphic DNA) 標誌進行選殖與分析,序列分析結果顯示其中兩個序列與逆轉位子 (retrotransposon) 的序列相關,另外二個序列則與玉米A染色體具有同源性。南方雜合分析 (Southern hybridization) 結果顯示此四個B染色體序列皆為重覆性序列。接著將四個B染色體專一性RAPD標誌轉換成SCAR (sequence characterized amplified region) 分子標誌,再以位於B染色體上不同斷裂點的B-10L易位染色體 (translocation),進行B染色體專一性SCAR分子標誌之實體定位,結果顯示四個B染色體專一性SCAR標誌皆位於異染色質區。螢光原位雜合分析 (fluorescence in situ hybridization) 顯示四個B染色體專一性RAPD序列在A染色體和B染色體中皆有訊號,在B染色體上的訊號集中於異染色質區域。最後應用此四個B染色體專一性SCAR標誌將帶CL-repeat的基因組片段定位於B染色體特定區域內。

The B-chromosome also called supernumerary chromosome is an extra or nonessential chromosomes that is not a member of the basic A chromosomes in individuals. The maize B-chromosome is a telocentric and it’s long arm consists of a proximal heterochromatic region, a proximal euchromatic region, a distal heterochromatic region and a distal ecchromatic tip. Isolation of sequences from the B-chromosome is always difficult, owing to that the DNA composition is similar between A and B chromosomes. Only three B-specific sequences (ZmBs, CL-repeat, and StarkB) are identified in maize until now. In this study, we cloned and analyzed four B-specific RAPD (random amplification of polymorphic DNA) markers. Sequences alignment showed that two sequences had homology with retrotransposons in maize, and others had homology to A chromosomes. Southern hybridization analysis indicated that the four B-specific RAPD markers were repetitive sequences. Subsequently, the four B-specific RAPD markers were converted into SCAR (sequence characterized amplified region) markers. Physical mapping of the four B-specific SCAR markers by B-10L translocations with different breakpoints in the B-chromosome, indicating that the four B-specific SCAR markers located on the distal heterochromatic region. By fluorescence in situ hybridization, signals of the four B-specific RAPD sequences were observed in A and B chromosomes, and concentrated on the distal heterochromatic region of the B-chromosome. Finally, these four B-specific SCAR markers were further applied to map the CL-repeat-carrying-genomic fragments onto definte regions of the B-chromosome.
其他識別: U0005-2608201315113500
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