蒙古牛BTG2启动子的克隆与初步功能分析

THE CLONING AND PRIMARY FUNCTIONAL ANALYSIS OF MONGOLIAN CATTLE BTG2 PROMOTER REGION

BTG2是新型细胞抗增殖因子BTG/Tob基因家族成员之一,在猪、牛中均报道存在品种和个体表达差异,但相关分子机理有待揭示。为阐明蒙古牛不同个体BTG2差异表达的分子机理,3个独立蒙古牛个体BTG2预测启动子序列(ATG起始密码子上游1766bp)被克隆、测序。序列比对发现,克隆序列与基因组组装BTG2启动子序列分别存在99.94%,98.64%,98.58%同源性,而且同源性最高的序列重组到哺乳动物表达载体pEGFP-N1后,由此获得的表达框架导入到HeLa细胞有受神经生长因子诱导表达的特性,该结果和已有报道一致。表明蒙古牛BTG2启动子序列被成功克隆,由此奠定了后续分离、鉴定该启动子上顺式元件和反式作用因子的基础。

BTG2 is one member of a novel anti- proliferation gene family,BTG/Tob family.Both in pig and cattle,it＇ s reported that BTG2 was differential expressed in breeds as well as in individuals.But the molecular mechanism underlying it remains to be revealed.In order to elucidate it,three presumed promoter regions of BTG2 origin from three individual Mongolian cattle were cloned and sequenced.The sequences blasting indicated that three cloned sequences shared 99.94%,98.64%,98.58%homologies with genome assembling Mongolian cattle BTG2 promoter region,respectively.Furthermore,when the most similar sequence recombinant into mammalian expression vector pEGFP- N1 and introduced into HeLa cells,the reporter protein enhanced green fluorescent protein（EGFP）were induced expression by never growth factor,which coherent with reported results.Together,it＇ s demonstrated that the promoter regions of Mongolian catde BTG2 were successfully cloned,which lay the foundation of further isolation and characterization of the cis- elements localizes in this promoter region and trans- factors which interaction with these cis- elements.