马红球菌酵母双杂交基因组文库的构建

Construction of Yeast Two-hybrid Genomic Library of Rhodococcus equi

目的构建马红球菌酵母双杂交基因组文库,为研究马红球菌的致病机理创造条件。方法将马红球菌的基因组DNA部分酶切,克隆到酵母双杂交系统的AD载体中,再检测文库的独立克隆数和插入序列的大小以确定文库的质量。结果获得1×106独立克隆数的基因组文库,插入片段的平均大小为2.3 kb。结论酵母双杂交基因组文库的构建,为进一步研究马红球菌的致病机制打下基础。

OBJECTIVE To construct two-hybrid genomic library of Rhodococcus equi, and study the pathogenic mechanism of R. equi. METHODS The genomic DNA of R. equi was partially digested and cloned into AD vector of two-hybrid system, the number of independent clone and the size of insert fragment were detected as the quality index of the library. RESULTS The independent clone of the library was 1 ×10^6 and the average size of insert fragment was 2.3 kb. CONCLUSIONS The pathogenic mechanism of R. equi is not clear and it is very important for this study to construct two-hybrid genomic library.