猪精浆蛋白基因PSP-Ⅰ和PSP-Ⅱ启动子序列克隆及生物信息学分析  

Cloning and bioinformatics analysis of promoter sequences of Porcine PSP-Ⅰ and PSP-Ⅱ

作　　者：周辉云[1] 李庆平[2] 吴晗[2] 何庆玲[2] 宋成义[2] 陈国宏[2] 

机构地区：[1]扬州大学生物科学与技术学院,扬州225009 [2]扬州大学动物科学与技术学院,扬州225009

出　　处：《生物信息学》2011年第1期50-53,59,共5页Chinese Journal of Bioinformatics

基　　金：农业部转基因生物新品种培育重大专项(2009ZX08010-019B和2008ZX08006-005)

摘　　要：分别用PCR方法扩增了1.7kbp和1.6kbp的猪PSP-Ⅰ和PSP-Ⅱ基因的启动子,并进行TA克隆,测序鉴定,测序结果用DNAstar程序与Genebank中的相应序列进行对比分析,结果显示与已发表序列的同源性分别为99.8%和96.3%。利用生物信息学的方法对克隆的1.7kbp和1.6kbp的猪PSP-Ⅰ和PSP-Ⅱ基因的启动子区进行了预测分析。PSP-Ⅰ和PSP-Ⅱ基因序列对比(mVista)分析发现,启动子0→-1000bp的保守性较高,其中0→-200bp核心启动子部分的序列同源性达到了100%,而-1000bp上游序列的保守性则较低。启动子的位置预测(Promoter Scan)结果显示,转录起始点上游200bp的区域为两基因的核心启动子位置。启动子区转录因子结合位点预测(TFSEARCH)发现,转录起始位点上游的1000bp区域内含有大量的顺式调控元件,并且得到了一系列潜在的转录因子结合位点。In this study,we amplified 1.7kbp and 1.6kbp promoters of seminal plasma protein Ⅰ and Ⅱ （PSP-Ⅰ and PSP-Ⅱ） genes respectively by PCR,and the PCR products were confirmed by sequencing positive TA clones.The sequencing results was submitted to align with the sequence deposited in Genbank by DNAstar program,the results showed the identities of cloned sequences of PSP-Ⅰ and PSP-Ⅱ promoters with published sequences were 99.8% and 96.3% respectively.Further bioinformatics analysis was applied to the cloned promoters of PSP-Ⅰ and PSP-Ⅱ.Sequence alignment between promoters of PSP-Ⅰ and PSP-Ⅱ by mVista revealed that region from 0 to -1000bp was highly conservative,and the identity of region from 0 to -200 was 100%,while the conservation in the upstream of -1000bp was low.The core promoters were located in the upstream of -200bp of transcription start site of PSP-Ⅰ and PSP-Ⅱ genes by online Promoter Scan progam.The online program （TFSEARCH） analysis revealed there were many cis elements in the region from -1000bp to transcription start site of PSP-Ⅰ and PSP-Ⅱ genes,a lot of potential transcription factor binding sites was identified as well.

关 键 词：生物信息学 PSP-Ⅰ PSP-Ⅱ 启动子 

分 类 号：Q518.2[生物学—生物化学]