摘要
以毛竹(Phyllostachys edulis)当年所发新叶为材料,用改良的CTAB法提取基因组DNA,通过超声波震断DNA,琼脂糖凝胶电泳回收大小为700 bp-2000 bp的片段,经T4 DNA聚合酶末端补平,与经过牛小肠碱性磷酸酶处理的pUC19载体连接,通过电转化转染受体菌XL-1Blue,构建了毛竹基因组文库。文库容量为2.8×107,通过PCR验证,插入片段大小为750 bp-2000 bp,插入效率为91%,符合基因组文库构建的要求,为毛竹特异功能基因的克隆和分子标记的研究奠定了基础。
The genomic DNA was extracted from the young leaves of Phyllostachys edulis with modified CTAB method. After the DNA was sonicated, the DNA with the fragments ranging from 700 bp to 2 000 bp was ligased into pUC19 vector/Sma I＋CIP, which was transformed into XL-1 Blue by electroporation transformation. Results showed that the library reached 2.8×10^7 in capacity and contained over 91% recombinant bacteria, in which inserts fragments ranging between 750 bp and 2 000 bp in sizes through PCR. The successful construction of the cDNA library has laid a base for the study on characterization and cloning of specific genes and molecular markers of Phyllostachys edulis.
出处
《世界竹藤通讯》
2008年第2期14-18,共5页
World Bamboo and Rattan
基金
国家自然基金项目(30771753)资助
关键词
毛竹
基因组文库
PCR
Phyllostachys edulis
genomic DNA library
PCR
