大鼠周围神经组织cDNA文库的构建

cDNA LIBRARY CONSTRUCTION OF RAT PERIPHERAL NERVE TISSUES

为了克隆周围神经组织中新近发现的营养因子、生长因子和诱向因子的基因，本研究以ＳＤ大鼠坐骨神经为材料，提取和纯化ｍＲＮＡ，逆转录合成ｃＤＮＡ双链，与Ａｄａｐｔｅｒ连接后，再与脱磷酸化处理的λｇｔ１１臂相连，体外包装，构建了大鼠周围神经组织的ｃＤＮＡ文库。经测定，该文库的容量＞１．５×１０６，重组百分比为９６．７％，ｃＤＮＡ片段为０．５～８Ｋｂ。经扩增后的滴度达１．４×１０１３ｐｆｕ／ｍｌ。该文库的建立为筛选和分离目的基因，为从基因水平深入研究神经系统发育、再生。

In order to obtain the cDNA clone of novel growth factors ,neurotrophic and neurotropic factors in peripheral nerve tissues ,rat sciatie nerves were homogenized followed by purification of mRNA ,synthesizing cDNA ,linking and finally ligation of the cDNA into λgt11 vector and in vitro packaging . The inserts length of cDNA was 0.5～8 kb .The capacity of cDNA library was 1.5×10 6 ,with 96.7% of recombinant rate .The titer of library was about 1.4×10 13 pfu/ml . The cDNA library from the rat peripheral nerve should be useful for further screening and isolating the target gene related to the mechanism of development ,regeneration and aging in molecular neurobiology .