菌落原位杂交法筛选特定菌种的方法初探  

Optimization of Colony in Situ Hybridization for Screening Specific Strains

作　　者：刘超兰[1] 王海英[1] 李文芳[1] 张文学[1,2] 赖登燡[3] 范鏖[3] 

机构地区：[1]四川大学轻纺与食品学院,四川成都610065 [2]四川大学锦江学院白酒学院,四川眉山620860 [3]四川水井坊股份有限公司,四川成都610036

出　　处：《酿酒科技》2013年第8期15-18,共4页Liquor-Making Science & Technology

基　　金：国家自然科学基金资助项目(31071585)

摘　　要：菌落原位杂交法是重组DNA技术中最常用的方法之一,但该方法存在着杂交背景复杂及杂交结果不稳定性等缺点,所以进一步优化杂交体系显得尤为迫切。本研究通过优化杂交体系,最终确定利用溶菌酶处理杂交膜和采用非严紧型洗脱方式洗膜,不仅使得杂交体系的杂交结信号清晰,而且杂交背景较干净。研究结果表明,在探针特异性好的基础上,采用优化的菌落原位杂交体系筛选特定菌种是可行的。Colony in situ Hybridization is one of the most widely-used methods in recombinant DNA technology. However, it is currently facing difficulties such as deep hybrid background and instability of results in hybridization. Therefore, the optimization of colony in situ hybridization system is necessary. In this research, through optimizing colony in situ hybridization system, we found that using lysozyme treatment system and the elution way of relaxation type for colony in situ hybridization, not only made positive dark dots distinguished clearly, but also resulted in clearer hybrid background. The results indicated that the use of colony in situ optimized hybridization system is feasible to screen specific strains if oligonucleotide probe is specific.

关 键 词：菌落原位杂交 寡核苷酸探针 地高辛 优化 

分 类 号：TS201.3[轻工技术与工程—食品科学]