Nogo66拮抗性多肽的筛选及其对脊髓损伤大鼠轴突再生的影响.pdfVIP

2010, 26 1 : 88~ 92 Chinese Journal ofNeuroanatomy Nogo-66 1* 1 2 1 1 1 1 陈金荣 , 石增立 , 杨成, 刘凤, 石磊 , 刘巍, 袁文丹 : 1. , 2. , 256603 : Nogo-66,: , 36, P R DNA , , 12, 12 Nogo-66 ELISA Nogo-66T 10, 1 d 50 g/50 l , HRP, BBB : 10, Nogo-66, NAPA, NAPB ELISA, NAPA Nogo-66 NAPB Nogo-66 P 0. 05 NAPA 46 , BBB P 0. 05 , HRP NAPA , : Nogo-66 NAPA NAPA Nogo-66; ; ; Selection of pepti e bin ing specifically to the Nogo-66 by ribosom e isplay an its infuence on axonal regeneration of injure spinal cor 1 1 2 1 1 1 1 Chen J inrong , Shi Z engli , Yang Cheng , L iu Feng , Shi Lei, L iu Wei , Yuan Wendan 1. Department of Pathophysilolgy, 2. Department of Anatomy, BinzhouM edical ollege, Binzhou, 256603, hina Abstract Objective:To select the peptide that bind specifically toNogo-66 by ribosome display and exam ine its effect to the regeneration of spinalcord. M etho s: 36 random-sequence oligonucleotides and prmi ersw ere designed and synthe- sized. A DNA random library was constructed by P R using a random-sequence oligonucleotides template. Then a cou- pled transcription/translation reaction in vitrow as proceeded by the use ofE. coli S30 ExtractSystem. The ribosome dis- play system including 12 random peptideswas used to screen the inhibitory peptide of Nogo-66. The affinity betweenNo- go-66 and the selected peptide was determ ined by ELISA methods. Spinal injury atT 10 levelwas established and selected peptides 50 g/50 l w ere injected through tailvein 1 d post- injury. HRP tracing methodw as used to observe the regen- eration of the axons and the recovery of nervous functionw as examined by BBB scoring. R esults: after ten rounds selec- tion, the sequence of the peptide binding specifically to the nogo-66 protein was obtained and corresponding peptides, NAPA andNAPB, were synthesized. ELISA results showed that the affinity betw een NAPA andNogo-66 was higher than that of NAPB P 0. 05 . Follow ing injection by tailvein, BBB scoring of rats in the expermi