【摘要】 利用RT-PCR方法扩增了猪ERK6基因编码区的序列,将扩增产物与pMD18-T载体连接,构建了重组质粒;重组质粒经PCR、酶切鉴定后进行测序;采用Northern杂交和半定量RT-PCR方法在猪不同部位骨骼肌中分析了ERK6基因转录本的个数、大小及组织表达谱。结果显示,所克隆的猪ERK6基因片段长1 113 bp,含有1个1 104 bp的开放阅读框(ORF),该ORF编码367个氨基酸;该基因与已报道的人ERK6基因核苷酸序列相似性为90%;猪ERK6基因在肌肉组织中只有一个转录本,大小约1.5 kb。此基因在骨骼肌中的表达量最高,心、子宫次之,卵巢最低;而在脂肪、胃、肝、脾、肺、肾、十二指肠和胰腺中未见表达。结果表明,猪ERK6基因与已报道的小鼠和人ERK6基因一样,主要在骨骼中特异表达。更多还原

【Abstract】 The novel porcine gene ERK6 was amplified by RT-PCR,and ligated into pMD18-T vector.The recombinant plasmid was identified by PCR amplification,digestion with restriction endonuclease and sequencing.The numbers and size of porcine ERK6 transcripts in skeletal muscle were obtained with Northern blotting and the expression levels in multiple tissues were detected by semi-quantitative RT-PCR. In results,the ERK6 cDNA was 1113bp in size and had an ORF of 1104bp encoding 367 amino acids.Homology of nucleotide sequence between the amplified porcine and the reported human ERK6 genes was 90%.The Northern blotting analysis showed that the gene had only one transcript which was 1.5kb approximately in size in skeletal muscle.It was highly expressed in skeletal muscle,less in heart and uterus,the lowest in ovary,but not detected in the other tissues including spleen,lung,kidney,fat,pancreas,small intestine,liver and stomach.The results showed that the porcine ERK6 gene expressed mainly in skeletal muscle as the same in mouse and human.更多还原