大鼠损伤神经组织中Tropic1808基因相关蛋白的亲和层析法纯化  

THE PURIFICATION OF TROPIC 1808 GENE ASSOCIATED PROTEIN IN INJURED SCIATIC NERVE OF RAT BY AFFINITY CHROMATOGRAPHY

作　　者：姚登兵[1] 王云丹[1] 陈雪[1] 丁斐[1] 

机构地区：[1]南通医学院江苏省神经再生重点实验室,南通226001

出　　处：《南通医学院学报》2002年第4期360-362,共3页ACTA Academiae Medicinae Nantong

基　　金：国家自然科学基金资助 (3 0 0 70 2 5 5 ) ;江苏省卫生厅资助 (HZ0 0 12 6)

摘　　要：目的 :纯化大鼠损伤坐骨神经组织中 Tropic180 8基因特异表达的蛋白。方法 :将 Tropic180 8基因重组蛋白的单克隆抗体与 CNBr- activated Sepharose 4 B交联 ,用亲和层析的方法 ,提取在大鼠损伤坐骨神经组织中Tropic180 8基因特异表达的蛋白 ,并将纯化的蛋白进行 SDS- PAGE分析。结果 :(1)经层析系统紫外检测 ,亲和层析后洗脱 ,在 2 80 nm波长处获得两个不同的紫外吸收峰。 (2 )亲和层析的洗脱液 ,经冷冻浓缩 ,SDS- PAGE分析 ,出现4 3.0 k Da、32 .0 k Da、31.0 k Da和 14 .4 k Da蛋白条带 ,其中在 32 .0 k Da的蛋白条带与预期的 Tropic180 8基因在组织中的编码蛋白基本相符 ,而 4 3.0 k Da、31.0 k Da、14 .4 k Da蛋白 ,预测为 Tropic 180 8基因在大鼠损伤坐骨神经组织中表达的相关蛋白。结论 :用亲和层析的方法 ,获得了大鼠损伤坐骨神经组织中 Tropic180 8基因特异表达的相关蛋白 。Objective:To purify the tropic 1808 gene expressed protein in injured sciatic nerve of rats.Methods:The monoclonal antibody of tropic 1808 gene fusion protein was coupled with CNBr-activated Sepharose 4B.By affinity chromatography,the tropic 1808 gene expressed protein was purified in injured sciatic nerve of rats.And then it was analyzed by SDS-PAGE.Results:(1)Two differentiated ultraviolet absorption peaks were obtained when the elution collected by affinity chromatography was analyzed at 280nm.(2)Four protein bands appeared at 43.0kDa,32.0kDa,31.0kDa and 14.4kDa respectively when the elution was analyzed by SDS-PAGE.And the protein band appearing at 32.0kDa was consistent with the expected.Conclusion:Purified tropic 1808 gene expressed protein in injured sciatic nerve of rats obtained by affinity chromatography. This study provides a basis for researching its structure and functions.

关 键 词：TROPIC 1808基因 大鼠 损伤坐骨神经 相关蛋白 单克隆抗体 亲和层析 SDS-PAGE 

分 类 号：R651.3[医药卫生—外科学]