为了研究香蕉束顶病毒与香蕉寄主致病的互作分子机制，本文报道利用Make Your Own＂Mate＆Plate＂ Library System试剂盒成功构建感染BBTV香蕉叶片的cDNA文库。通过改良CTAB法提取感染BBTV香蕉叶片的总RNA，采用SMART法反转录合成双链c...为了研究香蕉束顶病毒与香蕉寄主致病的互作分子机制，本文报道利用Make Your Own＂Mate＆Plate＂ Library System试剂盒成功构建感染BBTV香蕉叶片的cDNA文库。通过改良CTAB法提取感染BBTV香蕉叶片的总RNA，采用SMART法反转录合成双链cDNA，经靳，酶切并去除短片段之后，与经同样酶切的pGADT7-SfiI载体连接，利用电转法将重组载体转化到大肠杆菌宿主细胞中，获得初级cDNA文库，最后以初级文库100万克隆为基数扩增，得到扩增文库并提取质粒。结果得到库容量大于2．0X10。的初级文库，检测表明文库cDNA插入片段长度主要分布在700—2000bp，文库重组率为87．5％。结果表明，该文库质量较好，可用于后续酵母双杂交互作蛋白筛选试验，本研究为开展病毒与寄主互作的研究奠定基础。展开更多
RAPD assessment of genetic variations in 33 varieties of banana (Musa nana Lour.) was carried out. Eighteen primers were screened from 249 10 bp arbitrary primers, and a total of 192 DNA bands were amplified, among wh...RAPD assessment of genetic variations in 33 varieties of banana (Musa nana Lour.) was carried out. Eighteen primers were screened from 249 10 bp arbitrary primers, and a total of 192 DNA bands were amplified, among which 183 (95.31%) were polymorphic. The average number of DNA bands amplified by each primer was 10.67, and the average genetic distance among 33 varieties was 0.3412. Based on UPGMA cluster analysis of 192 DNA bands amplified by 18 primers a DNA molecular dendrogram was established for 33 varieties of banana in China, which divided the 33 varieties into four groups: group A, B, C and D. Group A included 20 varieties, group B included 5 varieties, group C included 2 varieties and group D included 6 varieties. Group A could be divided into 3 sub-groups: A1, A2 and A3. The molecular foundation of genetic diversity of banana was also explored.展开更多
文摘为了研究香蕉束顶病毒与香蕉寄主致病的互作分子机制，本文报道利用Make Your Own＂Mate＆Plate＂ Library System试剂盒成功构建感染BBTV香蕉叶片的cDNA文库。通过改良CTAB法提取感染BBTV香蕉叶片的总RNA，采用SMART法反转录合成双链cDNA，经靳，酶切并去除短片段之后，与经同样酶切的pGADT7-SfiI载体连接，利用电转法将重组载体转化到大肠杆菌宿主细胞中，获得初级cDNA文库，最后以初级文库100万克隆为基数扩增，得到扩增文库并提取质粒。结果得到库容量大于2．0X10。的初级文库，检测表明文库cDNA插入片段长度主要分布在700—2000bp，文库重组率为87．5％。结果表明，该文库质量较好，可用于后续酵母双杂交互作蛋白筛选试验，本研究为开展病毒与寄主互作的研究奠定基础。
文摘RAPD assessment of genetic variations in 33 varieties of banana (Musa nana Lour.) was carried out. Eighteen primers were screened from 249 10 bp arbitrary primers, and a total of 192 DNA bands were amplified, among which 183 (95.31%) were polymorphic. The average number of DNA bands amplified by each primer was 10.67, and the average genetic distance among 33 varieties was 0.3412. Based on UPGMA cluster analysis of 192 DNA bands amplified by 18 primers a DNA molecular dendrogram was established for 33 varieties of banana in China, which divided the 33 varieties into four groups: group A, B, C and D. Group A included 20 varieties, group B included 5 varieties, group C included 2 varieties and group D included 6 varieties. Group A could be divided into 3 sub-groups: A1, A2 and A3. The molecular foundation of genetic diversity of banana was also explored.
