利用PCR方法克隆了准噶尔雅罗鱼(Leuciscus merzbacheri)的β-actin基因启动子片段SZ21,大小是2398bp。对克隆的启动子序列进行了转录调控元件的生物信息学预测分析,同时,基于启动子中包含的开放阅读框和内含子序列,探讨了准噶尔雅罗鱼与鲤鱼(Cyprinus carpio)、草鱼(Ctenopharyngodon idella)、青鱼(Mylopharyngodon piceus)、团头鲂(Megalobrama amblycephala)、泥鳅(Misgurnus mizolepis)间的系统进化关系。结果显示,该启动子序列的3个核心启动子转录元件:CAAT-box、CArGmotif和TATA-box分别在转录起始位点(+1)上游的-89、-59、-26处,序列中还含有MEF2、SATB、CHRF、INRE、MTEN、E-box、RU49、ZBPF、CREB、Enhance region、CEBP位点等多种转录调控元件。在剪接体内含子中,剪接位点遵循GT…AG法则。启动子SZ21序列含有3个内含子和155个氨基酸。内含子1、内含子2、内含子3的系统发育分析表明,团头鲂与草鱼和青鱼的亲缘关系要比与准噶尔雅罗鱼的更近一些,这与传统分类中的亲缘关系显示不一致,其原因尚需探讨。
Through PCR amplification, the β-actin gene promoter fragment SZ21 (2 398 bp) of Leuciscus merzbacheri was obtained. We forecasted its transcriptional regulation elements using bioinformatics method. In view of its ORF and introns sequence, the system development was analyzed between L. merzbacheri and other closely related fish species. Three core promoter transcriptional elements were situated in the transcriptional start site( + 1 ) upper - 89, - 59, - 26, including the CAAT-box, CArG motif, TATA-box. Results of putative transcription binding sites analysis of the promoters revealed the presence of several critical transcription binding sites such as the MEF2, SATB, CHRF, INRE, MTEN, E-box, RU49,ZBPF, CREB, Enhance region and CEBP in the further promoter. The splice sites abided by GT… AG rule in the spliceesome introns. The promoter SZ21 sequence included the intron 1, intron 2, intron 3, and 155 amino acids. The system development analysis of three introns showed that Megalobrama amblycephala had more close relationship to Ctenopharyngodon idella and Mylopharyngodon piceus than L. merzbacheri. This is not consistent with the traditional classification, and the reason needs further investigation.