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郑鹏飞,刁晓平,李赞东,杜俊俏.2016.蚯蚓Sox2基因的克隆及其在再生进程中的表达.动物学杂志,51(1):95-102.
蚯蚓Sox2基因的克隆及其在再生进程中的表达
Gene Cloning and Expression of Sox 2 during Tail Regeneration of Earthworm Eisenia foetida
投稿时间:2015-02-10  修订日期:2015-10-30
DOI:DOI: 10.13859/j.cjz.201601011
中文关键词:  赤子爱胜蚓  Sox2  克隆  再生  RACE
英文关键词:Eisenia foetida  Sox2  Clone  Regeneration  RACE
基金项目:海南省普通高等学校研究生创新科研课题 基金编号:No. S201305
作者单位E-mail
郑鹏飞 海南大学 zhengpengfei122@126.com 
刁晓平 海南大学 diaoxip@hainu.edu.cn 
李赞东 中国农业大学  
杜俊俏 海南大学  
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中文摘要:
      为了解Sox2基因在赤子爱胜蚓(Eisenia foetida)再生进程中的作用,本研究采用cDNA末端快速扩增(RACE)方法成功克隆并得到了完整的赤子爱胜蚓Sox2基因序列(GenBank登录号:KP637161),其cDNA序列全长2 354 bp,其中包括367 bp的5′端非翻译区,844 bp的3′端非翻译区和编码380个氨基酸残基的1 143 bp开放阅读框。通过实时荧光定量PCR检测了Sox2基因在发育成熟的赤子爱胜蚓不同体段(头部、环带和尾部)以及在尾部体段再生进程中的表达特征。结果显示:Sox2在不同体段中(头部、环带、尾部)的表达差异不显著(P>0.05)。在尾部断肢后再生进程中,随着时间推移,Sox2的表达量明显上调,其中,截断后12 h,Sox2基因表达量达到峰值,是截断初期(0 h)的22倍。研究结果表明,Sox2基因可能与蚯蚓的再生进程有关。
英文摘要:
      In order to elucidate the role of Sox2 in the earthworm Eisenia foetida during regeneration.,the full length cDNA of Sox2 was firstly cloned from earthworm by using rapid amplification of cDNA ends (RACE) approach. The expression level of Sox2 in different body segments (head, clitellum and tail) of mature earthworm and during tail segments regeneration at different time point was detected by quantitative real-time PCR. The data was analyzed by the method of Delta-Delta Ct. RACE results showed that the full-length of cDNA sequence is 2 354 bp (GenBank Accession number:KP637161), including 367 bp of 5'-UTR, 844 bp of 3 '-UTR and 1 143 bp ORF (Fig. 1 and 2). Phylogenetic analysis showed Sox2 in Eisenia foetida was conservative in phylogeny with other species (Fig. 3).The q-PCR results showed that the Sox2 expression transcripts were not significantly difference in head, clitellum and tail body segments (P>0.05) (Fig. 4a). The expression level of Sox2 in the tail was obviously up-regulated during regeneration, and reached the highest level at 12 h after amputation with 22 times compared with that in the beginning of amputation (Fig. 4b).These results indicated that the Sox2 gene may be associated with the regeneration in earthworm. Our research provided new evidence for further understanding of the molecular mechanisms of the regeneration.
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