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目的 构建福氏 2a志贺氏菌 3 0 1株染色质DNA基因文库 ,为下一步全基因组序列测定打下基础。方法 Murry法提取福氏 2a志贺氏菌染色质DNA ,采用鸟枪法策略 ,分别经 8种限制性内切酶消化后克隆至载体质粒中 ,并经双脱氧链末端终止法测序后计算重组质粒重复率。结果 Murry法提取福氏 2a志贺氏菌染色质DNA大小为 3 0Kb以上 ,共构建 86 3 2个重组质粒 ,质粒重复率为 1 %。结论 本研究在国际上首次成功构建了福氏 2a志贺氏菌株染色质DNA基因文库 ,文库全长为福氏 2a志贺氏染色质的 6倍 ,已足够全基因组序列测定所需。
Objective To construct a library of 301 chromosomal DNA genes of Shigella flexneri that lays the foundation for the next step genome-wide sequencing. Methods Chromosome DNA of Shigella flexneri 2a was extracted by Murry method. After being digested with 8 kinds of restriction enzymes, it was cloned into plasmid vector and sequenced by dideoxy chain termination method to calculate the recombinant plasmid Repeat rate. Results Murry extraction of Shigella flexneri 2a Chlamydia chromatin DNA size of more than 30K, a total of 86 3 2 recombinant plasmids were constructed, the plasmid repetition rate was 1%. Conclusions This study was the first successful in the world to construct the Cholera DNA library of Shigella flexneri 2a strain. The full-length cDNA library of Chlamydia pubescens was 6 times of that of Cholera gerbil 2a, which was enough for genome-wide sequence analysis.