摘要
利用显微切割技术，分离了黑麦（ＳｅｃａｌｅｃｅｒｅａｌＬ．）１０个Ｂ染色体短臂端部片段，并利用寡核苷酸引物（ＣＣＣＴＡＡＡ）３及新建立的二级单引物序列ＰＣＲ扩增法，扩增了黑麦Ｂ染色体端粒相关序列。染色体原位杂交实验将ＰＣＲ产物定位于Ｂ染色体短臂末端，多数Ａ染色体末端也显示清晰的杂交信号。部分ＰＣＲ产物克隆到ｐＵＣ１９载体中，对其中一个克隆子ｐｐ３的序列分析结果表明，它与玉米亚端部克隆子ｐＢＦ２６６部分区域的同源性为９２％。就目前资料检索，黑麦、玉米端粒相关序列具有高度同源性还未见报道。
The short arm terminal segments of 10 rye (Secale cereal L.) B chromosomes were isolated by means of needle microdissection technology. A two step single primer PCR method was used to amplify the telomere associated sequences of rye B chromosomes. The PCR products were located in the terminal region of the short arm of rye B chromosomes by chromosome in situ hybridization, and most A chromosomes also showed clear signal dots. Some PCR products were cloned in pUC19 vector and one clone pp3 was sequenced. Sequence analysis demonstrated that it has high similarity with the maize subtelomeric clone pBF266. Further utility of this microdissection PCR system in construction of high density RFLP map was discussed.
基金
国家自然科学基金
关键词
黑麦
B染色体
显微切割
端粒相关序列
Rye B chromosomes, Microdissection, Telomere associated sequences
