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標題: Physical mapping of RFLP markers and translocations on maize chromosome 8
玉米第八對染色體之易位染色體斷裂點與 RFLP 標誌的實體定位
作者: 謝淳芷
Hsieh, Chun-Chih
關鍵字: 第八對染色體;斷裂點;實體定位
出版社: 分子生物學研究所
玉米 r-X1 缺失是第十條染色體長臂上一小段區域缺失。它會引起低頻率的母本染色體整條丟掉外,也會導致母本染色體斷裂,產生尖端缺失的染色體。整條染色體缺失謂之完全單染體 (complete monosome,CM),而尖端缺失謂之部分單染體 (partial monosome, PM)。
本實驗室多年來利用這些不同程度缺失的PM標定RFLP標誌。但先前的系統不能取得斷裂點分布在整條染色體臂的PM。為了解決這個問題,本論文結合r-X1 缺失與A-A易位染色體,把含適當篩選基因的染色體片段接到特定的染色體臂尖端,然後再利用該基因篩選PM。
第八條染色體短臂 (8S) 含很少基因,更沒有可供篩選PM 的基因。故本實驗擬以A-A易位染色體進行篩選該臂PM,藉以從事RFLP標誌的實體定位。研究中所使用的材料有二: 一為T8-9b,是8S接上部分第九條染色體的長臂 (9L); 二為T6-8b,即8S接上部分第六條染色體的長臂 (6L)。本論文的目的在利用這兩個易位染色體配合r-X1缺失以產生CM及PM,一方面定位T8-9b及T6-8b的斷裂點,一方面標定8S上的RFLP 標誌。
此系統在T8-9b方面共得到6棵8S PM、 5棵8L PM 及20棵T8-9b CM,這些材料將T8-9b在8S的斷裂點標定於bnl7.08與bnl9.08間,而9L上的斷裂點定位在bnl14.28及umc140之間。至於T6-8b方面,得到3棵8S PM、 5棵8L PM 及13棵T6-8b CM,在6L上的斷裂點位於umc59a的外側,但8S上的斷裂位置則在bnl9.08的內側。另外,標定RFLP 標誌方面,將bnl9.08定位於8L PM之斷裂點的內側,而csu54b和bnl14.28定位於兩8S PM的外側,並將此兩8S PM的斷裂點定於bnl14.28與bnl9.08之間。

The r-X1 deletion is an interstitial deficiency located on the long arm of chromosome 10. It can induce not only the formation of monosome (or complete monosome, CM) but also break any chromosome arm. The resulting centromere-carrying chromosome is terminally deficient (PM).
In the past, our laboratory used PM to map RFLP markers. The previous system does not allow screening PM with breakpoints spreading over the entire chromosome because of shortage of proper selective genes in most chromosome arms. In this thesis, a new approach was adopted to overcome this shortcoming. It involves using the r-X1 deletion in conjunction with an A-A translocation carrying a desirable selection marker and a breakpoint near the terminal position of the chromosome arm of interest.
The short arm of chromosome 8 (8S) carries very few genes, none of which can be used as selection marker for PM isolation. We are attempting to use A-A translocations carrying appropriate selection marker to isolate PMs on this arm and used them to map physically RFLP marker. Two translocations were used in this study: T8-9b carrying 8S linked to the terminal half of the long arm of chromosome 9 (9L) and T6-8b containing 8S attached to the terminal half of the long arm of chromosome 6 (6L). The objective of this thesis is using the two translocations together with the r-X1 to produce CMs and PMs for mapping the breakpoints of the translocations and RFLP markers on 8S.
The use of T8-9b produces six 8S PMs, five 8L PMs and 12 89 CMs,which enable determination of the breakpoint of T8-9b in the region between bnl7.08 and bnl9.08 on 8S and between bnl14.28 and umc140 on 9L. On the other hand, the employment of T6-8b resulted in three 8S PMs, five 8L PMs and 13 68 CMs. The translocation breakpoint was partially analyzed, being distal to umc59a on 6L but proximal to bnl9.08 on 8S.
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