遗传 ›› 2000, Vol. 22 ›› Issue (4): 197-200.
• 论文 • 下一篇
李连城1;何聪芬1;2;徐琼芳1;马有志1;辛志勇1 LI Lian-cheng1;HE Cong-fen1;2;XU Qiong-fang1;MA You-zhi1;XIN-yong1
摘要: 本研究利用微细玻璃针法从减数分裂中期I的花粉母细胞中分离回收了携带小麦抗黄矮病基因的中间偃麦草染色体2Ai-2。将目标染色体放入装有蛋白酶K消化液的0.5ml小离心管中,用Sau3AI酶切,在DNA片段的末端连接接头后,进行 PCR扩增。扩增产物经琼脂糖凝胶电泳分析,DNA片段的大小在150~3000bp之间,而大部分集中在200~1500bp之间。
Abstract:A simple method was used to adapt refined needles for the collection of Th.intermedium 2Ai-2 chromosome carrying BYDV (barley yellow dwarf virus) resistance gene from meiosis-metaphase spreads .The aimed chromosome was put into a 0.5ml Eppendorf tube,deproteinized with proteinase K,digested with Sau3AI,and link-adaptors were ligated to the ends of the DNA fragments.After amplification by PCR,size distributions of the PCR products were analyzed by agarose gel electrophoresis and smears of DNA were revealed that the size ranged from 150bp to 3000bp with predominant fragments at about 200~1500bp.