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鲤(Cyprinus carpio)胰岛素样生长因子2b(insulin-like growth factor 2b,IGF2b)基因为鲤IGF基因家族重要的成员,已有的研究表明5’侧翼区序列对其功能的发挥有重要作用。因此,采用基因组步移法获取5’侧翼区序列,通过亚硫酸盐修饰的方法分析不同品种该区域GC富集区的甲基化状况。本文成功从鲤基因组DNA中获得695 bp的IGF2b 5’侧翼区序列。已获取的IGF2b 5’侧翼区序列经过TFSEARCH分析后发现了多个转录因子结合位点和TATA框,通过BLAST比对发现,鲤IGF2b 5’侧翼区DNA序列与斑马鱼(Danio rerio)IGF2b基因相比,相似性为87%。检测该区域富含GC的序列17个位点,发现建鲤(C.c.var.jian)有2个个体的3个位点出现甲基化,而黄河鲤(C.c.haematopterus)只有1个个体1个位点出现甲基化,这说明2个品种鲤在这个区域出现甲基化修饰的几率低,也验证了这2品种在该基因该区域GC富集区是稳定的。
Common carp (Cyprinus carpio) insulin-like growth factor 2b (IGF2b) gene is an important member of the common carp IGF gene family. Some studies have shown that the 5 ’flanking region sequence plays an important role in its function. Therefore, the 5 ’flanking region sequence was obtained by genome walking method and the methylation status of GC-rich regions in different species was analyzed by sulfite modification method. In this paper, 695 bp 5’-flanking region of IGF2b was successfully obtained from common carp genomic DNA. The obtained 5 ’flanking region of IGF2b showed multiple transcription factor binding sites and TATA box after TFSEARCH analysis. BLAST analysis showed that the 5’ flanking region of IGF2b shared the same sequence with the Danio rerio IGF2b gene The similarity is 87%. The results showed that methylation was detected in 3 loci of two individuals in Ccvar.jian and only one locus in Cchaematopterus Point methylation, indicating that the two species of common carp in this region there is a low probability of methylation modification, also verified these two varieties of the gene in this area GC-rich area is stable.