摘要
综述了启动子序列克隆和功能研究方法的研究进展。启动子克隆的主要方法有基因组文库筛选法、探针载体筛选法、常规PCR法、反向PCR法、锅柄PCR法、序列特异性引物PCR法、热不对称交错PCR法和Y型接头扩增法。其中,热不对称交错PCR法因操作简便、特异性较高而最有效。启动子功能分析方法有生物信息学分析法和实验分析法,前者主要依托数据库初步预测启动子序列;后者确定启动子的顺式元件及其功能,具体策略有点突变分析、凝胶阻滞分析、瞬间表达分析、转化分析和酵母单杂交分析。可为启动子功能的研究提供参考。
In this paper, the research advances in the methods of the cloning and function-analyzing of promoters were reviewed. The main methods used for promoter cloning are genomic library-screening, probe vector-screening, conventional polymerase chain reaction （PCR）, inverse PCR （I -PCR）, panhandle PCR （P- PCR）, sequence specific primers PCR （SSP- PCR）, thermal asymmetric interlaced PCR （ TAIL - PCR）, and Y - shaped adaptor dependent extension （ YADE）, of which TAIL -PCR is the most effective one, because it is easy to operate and with high specificity. Approaches of promoter function-analysis include bioinformatical and experimental analysis. The bioinformatical analysis relies on database searching to predict promoter sequences. The experimental analysis can determine the cis-acting elements and their functions. The concrete analytical strategies included point mutation, gel retardation, transient expression, transformation and yeast one-hybridization. This review can provide a reference for the function-researching of promoters.
出处
《云南农业大学学报（自然科学版）》
CAS
CSCD
北大核心
2010年第2期283-290,共8页
Journal of Yunnan Agricultural University:Natural Science
基金
云南省科技厅基金项目(2006C0030Q)
云南农业大学博士启动基金项目(A2002096)
关键词
启动子
序列克隆
功能分析
方法
promoter
sequence-cloning
function-analyzing
methods
