Sox8基因HMG盒区内含子剪接位点分析

Sox基因参与广泛的发育调控过程.为了确定Sox8基因HMG盒区内含子的大小及剪接位点,通过计算机分析本室克隆得到的泥鳅Sox8基因包括HMG盒区在内的一段基因组序列,推测在HMG盒区可能存在一个内含子,进一步通过RT-PCR方法,克隆泥鳅Sox8基因HMG盒区cDNA片段,与基因组序列比较分析,确认在泥鳅Sox8基因的HMG盒区存在一个内含子,并确定了该内含子的序列及剪切位点.同时,比较分析了人和泥鳅Sox8在HMG盒区的内含子的剪切位点.结果显示,Sox8基因HMG盒区内含子剪切位点在进化上是保守的。 Abstract:Sox genes have diverse roles in developing processes,it was inferred there may be one intron in the HMG box by analyzing the genomic DNA sequence of the paramisgurnus dabryanus Sox8 gene.RT-PCR was used to verify the intron and its splicing site.First strand of the cDNA reverse-transcribed from liver RNA was amplified using the primers flanking the HMG box.RT-PCR products were cloned into the pUC18 plasmid and sequenced.The eomparison between the cDNA and genomic DNA sequence revealed the splicing site of the intron existing in the HMG-box of the paramisgurnus dabryanus Sox8 gene.We also compared the splicing site of the intron in the HMG-box of Sox8 gene between paramisgurnus dabryanus and human.These results suggest that the splicing site in the HMG-box of Sox8 gene was conserved.

Analysis of the Splicing Site in the HMG-box of Sox8 Gene

Sox genes have diverse roles in developing processes.it was inferred there may be one intron in the HMG box by analyzing the genomic DNA sequence of the paramisgurnus dabryanus Sox8 gene.RT PCR was used to verify the intron and its splicing site.First strand of the cDNA reverse transcribed from liver RNA was amplified using the primers flanking the HMG box.RT PCR products were cloned into the pUC18 plasmid and sequenced.The comparison between the cDNA and genomic DNA sequence revealed the splicing site of the intron existing in the HMG box of the paramisgurnus dabryanus Sox8 gene.We also compared the splicing site of the intron in the HMG box of Sox8 gene between paramisgurnus dabryanus and human.These results suggest that the splicing site in the HMG box of Sox8 gene was conserved.