摘要
目的研究内含子和5＇非翻译区对血小板生成素(TPO)基因表达的影响.方法以中国人胎肝mRNA为模板,应用RT-PCR和长距离PCR(LD-PCR)技术扩增了约1.1kb的全长人TPO cDNA.6.2 kb的TPO基因组DNA和TPO基因组中的内含子I,内含子Ⅱ～Ⅳ和内含子Ⅴ.结果全序列分析表明,全部的编码序列、所有的内含子/外显子剪切部位序列同已知一致,个别差异发生在内含子中.结论通过PCR扩增技术,成功地从中国人胎肝组织中克隆了全长人TPO cDNA、TPO基因组DNA及其全部的内含子.
Objective To studytheroleof thrombopoietin(TPO)gene5'untranslatedregion(UTR)andtheintronin TPO expressionregulation.Methods WiththemRNAderivedfromChinesehumanfetalliveras thetemplate,full-lengthTPO cDNA(approximately1.1kb)andTPOgenomicDNA(6.2kb)alongwithallthe intronsof TPOgenewereisolatedby PCR andlong-distancePCRtechniquesfromChinesefetallivertissue.Result Sequencinganalysisindicatedthatallthecoding sequencesandtheexon/intronsplicesiteswereconsistentwiththeresultspreviouslyreportedby Sohmaet al exceptforonlya fewnucleotidesintheintronof TPOgene.Conclusion Wehavesuccessfullyclonedfull-lengthTPOcDNA,TPOgenomic DNAandalltheintronsof TPOgenefromChinesefetallivertissue.
出处
《第一军医大学学报》
CSCD
北大核心
2001年第12期881-884,共4页
Journal of First Military Medical University
基金
国家科委"1035"重大项目(96-901-01-009)
广东省自然科学基金重点项目(963011)
