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標題: Establishment and application of CL-repeat display markers in maize B chromosome
作者: 簡鈺倫
Yu-Lun Chien
關鍵字: maize;B-chromosome;display technique;CL-repeat;玉米;B染色體;顯示技術;CL-repeat
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Regional control of nondisjunction of the B chromosome in maize. Genetics 90: 613-627. Lin, B.Y. 1979. Two new B-10 translocations involved in the control of nondisjunction of the B chromosome in maize. Genetics 92: 931-945. Lin, B.Y. and H.P. Chou. 1997. Physical mapping of four RAPDs in the B chromosome of maize. Theor. Appl. Genet. 94: 534-538. Longley, A.E. 1927. Supernumerary chromosomes in Zea mays. J. Agric. Res. 35: 769-784. Masonbrink, R.E. and J.A. Birchler. 2010. Sporophytic nondisjunction of the maize B chromosome at high copy numbers. J. Genet. Genom. 37: 79-84. McClintock, B. 1933. The association of non-homologous parts of chromosomes in the mid-prophase of meiosis in Zea mays.Z. Zellf. Mik. Anat. 19: 191-237. Peng, S.F., Y.P. Lin and B.Y. Lin. 2005. Characterization of AFLP sequences from regions of maize B chromosome defined by 12 B-10L translocations. Genetics 169: 375-388. Randolph, L.F. 1928. Types of supernumerary chromosomes in maize. Anat. Rec. 41:102. 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為了獲得大量玉米B染色體專一性的分子標誌,利用B染色體專一性序列CL-repeat廣泛分佈在B染色體的特性,並參考鳶尾逆轉位子顯示技術,建立玉米B染色體CL-repeat顯示技術。總共獲得26個B染色體專一性標誌,並利用5個B-10L易位染色體進行標誌定位,再利用13個位於異染色質區的B-10L易位染色體進行相互定位,藉以確定易位染色體斷裂點的位置並精細定位各個CL-repeat顯示標誌。將26個標誌進行選殖與序列分析,依序列特性可分成CL-repeat序列標誌、B染色體中節序列標誌、同時帶有CL-repeat與B染色體中節序列標誌及未知序列標誌此4大類。接著將1個B染色體中節序列標誌、2個同時帶有CL-repeat與B染色體中節序列標誌及2個未知序列標誌轉換成SCAR (sequence characterized amplified region)標誌,並利用B-10L易位染色體進行定位,其中帶B染色體中節序列之3i-08-SCAR標誌出現在非預期的區域,將不同區域的標誌進行選殖與定序,並製成無根樹演化圖,以探討玉米B染色體中節序列在中節與遠端異染色質區的差異。另外,以限制酵素HindIII進行CL-repeat顯示技術,期望獲得更多不同的B染色體專一性的CL-repeat顯示標誌。

For isolating a large number of maize B-chromosome specific molecular markers, we used the B-chromosome specific sequence, CL-repeat which are widely distributed on the B-chromosome, and referred to the IRRE (iris retroelement) retrotransposon display technique to establish the CL-repeat display technique of maize B-chromosome. A total of 26 B-chromosome specific markers were obtained and mapped by five B-10L translocations. Additonal 13 B-10L translocations with breakpoints on the distal heterochromatic region were used to verify breakpoints of these translocations and to finely map each CL-repeat display marker. All of the 26 markers were cloned and sequenced. On the basis of sequence characteristics, those sequence could be divide into four categories: markers with CL-repeat sequence, markers with B-centromere sequence, markers with CL-repeat and B-centromere sequences, and markers with unknown sequence. Subsequently, one marker with B-centromere sequence, two markers with CL-repeat and B-centromere sequences, and two markers with unknown sequence were selected to convert into SCAR (sequence characterized amplified region) markers, and mapped by B-10L translocations. Among these five markers, the 3i-08-SCAR marker with B-centromere sequence were mapped to unexpected regions. Those unexpected SCAR markers were cloned and sequenced from different regions, and used to constructed an unrooted evolution tree for exploring the differentiation of maize B-centromere sequences between the B-centromere and the distal heterochromatin region. Furthermore, we used restriction enzyme HindIII to perform the CL-repeat display technique and hoped to get more different B-chromosome specific CL-repeat display markers.
其他識別: U0005-2702201412134200
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