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c DNA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(Gen Bank登录号:KP637161),其c DNA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。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。
In order to elucidate the role of Sox2 during regeneration in the earthworm Eisenia foetida, the full length c DNA of Sox2 was firstly cloned by using rapid amplification of c DNA ends（RACE） approach. The expression level of Sox2 in different body segments（head, clitellum and tail） of mature earthworm and during tail regeneration at different time points was detected by quantitative real-time PCR. The data were analyzed by the method of delta-delta Ct. RACE results showed that the full-length of c DNA sequence was 2 354 bp（Gen Bank 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 that Sox2 in Eisenia foetida was conservative in phylogeny compared with other species（Fig. 3）. The q-PCR results showed that Sox2 expression transcripts were not significantly different in head, clitellum and tail body segments（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, 22 times higher 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 has provided new evidence for further understanding of the molecular mechanisms of the regeneration.
出处
《动物学杂志》
CAS
CSCD
北大核心
2016年第1期95-102,共8页
Chinese Journal of Zoology
基金
海南省普通高等学校研究生创新科研课题(No.S201305)
