生殖医学杂志2011，Vol.20Issue(6)：501-506,6.DOI:10.3969/j.issn.1004-3845.2011.06.016
真核表达CatSper1用于筛选有效siRNA的体外实验研究
Study of seeking effective siRNA targeting mouse CatSper1 by using a pEGFP-N1-CatSper1 eukaryotic expression vector
摘要
Abstract
Objective: To find effective siRNA targeting mouse CatSperl mRNA that was transcribed by a recombinant plasmid pEGFP-N1-CatSperl.Methods: The full length of mouse CatSperl cDNA was cloned using recombinant PCR and integrated into a pEGFP-Nl plasmid (pEGFP-Nl-CatSperl). Three siRNAs were predicted with the online bioinformatics analysis and synthesized. The synthesized siRNAs were co-transfected into the N2a cells with the pEGFP-Nl-CatSperl plasmid. The interferential effect of siRNAs was determined by EGFP fluorescence intensity, quantitative PCR and Western blot.Results: The full length of mouse CatSperl cDNA was successfully cloned into the pEGFP-Nl plasmid. CatSperl mRNA and protein expressions were substantially inhibited by the siRNA located on the 3' of CatSperl mRNA, while all the three siRNAs showed interferential effects. Negative control siRNA did not cause changes in CatSperl mRNA and protein expressions.Conclusion: The three effective siRNAs targeting CatSperl were identified and could be used for the study of the function and contraceptive potential of CatSperl.关键词
CatSper1基因/RNA干扰/共转染Key words
CatSperl gene: RNAi/Co-transfection.引用本文复制引用
梅小琴,李红钢,熊承良..真核表达CatSper1用于筛选有效siRNA的体外实验研究[J].生殖医学杂志,2011,20(6):501-506,6.基金项目
国家自然科学基金项目(30770814),湖北省计划生育委员会科研基金 （30770814）
