高质量水稻基因组文库构建的策略和方法  

Strategy and Methods of Construction High Quality Rice Genomic Library

作　　者：梁卫红[1] 刘一鸣[2] 吴乃虎[2] 

机构地区：[1]河南师范大学生命科学学院,河南新乡453007 [2]中国科学院遗传与发育生物学研究所

出　　处：《河南师范大学学报（自然科学版）》2006年第2期177-179,共3页Journal of Henan Normal University(Natural Science Edition)

基　　金：国家重大基础研究发展计划(2001CB1088);国家"863"计划(2002AA224061);河南省高校青年骨干教师资助计划(521697);河南师范大学青年科学基金(521911)

摘　　要：采用改良的CTAB法提取水稻基因组DNA,经Sau3AI部分酶切的产物经透析袋电洗脱法回收、正丁醇浓缩后,以1∶1的摩尔数比和载体臂连接.包装后测定,文库滴度达到106 pfu/ml;随机酶切重组克隆显示,插入片段达到预期的15～23 kb之间,提示该方法适于植物材料高质量基因组文库的构建,有利于分离完整的基因序列及其调控区.Rice genomic DNA had been isolated by improved CTAB method, and the Sau3AI partial-digestion products were recovered by electric elution with dialysis bag. After concentrated by n-butyl alcohol, the purified partial digestion products were ligated with vector arms in the 1 ： 1 ratios. After packaging, the titer reached to 10^8 pfu/ml. Quality detection showed that the DNA inserts were between 15～23 kb as expect, suggest that the method was appropriate for constructing plant genomic library, and was helpful to isolate full-length gene and its regulation sequences.

关 键 词：基因组DNA 部分酶切 电洗脱 滴度 

分 类 号：Q78[生物学—分子生物学]