张芳芳, 马岚. 小鼠胚胎干细胞R1表面特异结合多肽的筛选[J]. 云南大学学报(自然科学版), 2012, 34(S1): 85-92.
引用本文: 张芳芳, 马岚. 小鼠胚胎干细胞R1表面特异结合多肽的筛选[J]. 云南大学学报(自然科学版), 2012, 34(S1): 85-92.
Screening of peptides binding specificially to surface moleculesof mouse embryonic stem cells R1[J]. Journal of Yunnan University: Natural Sciences Edition, 2012, 34(S1): 85-92.
Citation: Screening of peptides binding specificially to surface moleculesof mouse embryonic stem cells R1[J]. Journal of Yunnan University: Natural Sciences Edition, 2012, 34(S1): 85-92.

小鼠胚胎干细胞R1表面特异结合多肽的筛选

Screening of peptides binding specificially to surface moleculesof mouse embryonic stem cells R1

  • 摘要: 胚胎干细胞是从哺乳动物胚胎发育早期的囊胚的内细胞团内分离出来的一类细胞,具有自我更新和全能性的基本特征.作者利用M13 噬菌体展示技术筛选与未分化的小鼠胚胎干细胞R1表面特异结合的多肽.实验利用未分化的小鼠胚胎干细胞为筛选靶细胞,分化的小鼠胚胎干细胞为吸附细胞,进行3轮的消减筛选,经细胞ELISA 鉴定,噬菌体DNA 测序和多肽序列分析,得到13条能与小鼠胚胎干细胞特异结合的多肽.通过BLASTP 比对发现有11个体内的同源蛋白,为进一步研究小鼠胚胎干细胞表面分子提供研究基础.

     

    Abstract: Embryonic stem cells (ESCs) are cells derived from the inner cell mass of the blastocyst of early-stage embryo,ESCs are distinguished by two distinctive properties:self-renewal and pluripotency.Our study aimed to screen and identify peptides that specifically bind to surface molecules of mouse ESC R1 using the phage display technology to provide foundation for identifying new R1 cell surface markers.During the screening process,only peptides that could bind to R1 surface but not differentiated cells surface were considered potential target peptides.After three rounds of screening,phages specifically binding to R1 surface were obtained and the corresponding peptides’ sequences were determined through analyses of the inserted DNA sequences.Then we performed ELISA to validate our results using differentiated cells and MEF cells as controls and confirmed the binding specificity of these peptides to R1.Finally,we analyzed the homology between our potential target peptides with known proteins and discussed possible corresponding protein identities,which provided important cues for identification of R1 surface markers.

     

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