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为克隆神经损伤及再生过程中出现的诱向因子和营养因子基因，以ＳＤ大鼠手术损伤的坐骨神经组织为材料，提取ｍＲＮＡ，逆转录合成ｃＤＮＡ，以λｇｔ１１为载体，构建了一个ｃＤＮＡ文库。经测定，该文库的容量大于１．５×１０６，重组百分率为９６．７％，插入片段长度分布在０．５～８＼＾０ｋｂ之间。扩增后的滴度达１．４／１０１２ｐｆｕ／ｍｌ。以该文库ＤＮＡ为模板，运用ＰＣＲ方法扩增出３６０ｂｐ的ＮＧＦ和６１６ｂｐ的ＣＮＴＦ阅读框架片段。以地高辛标记的ＣＮＴＦ探针，从文库中筛选出ＣＮＴＦ克隆。经ＤＮＡ测序证实确为ＮＧＦ和ＣＮＴＦｃＤＮＡ全序列。根据已获得的６５ｋＤ化学诱向因子抗体，应用ＡＢＣ法筛选文库，获得了阳性克隆，其插入片段为１．１ｋｂ左右。经Ｇｅｎｂａｎｋ查询证实为一新序列。该文库的建立为筛选和分离目的基因，从基因水平深入研究神经再生、发育及退行性变的机制奠定了基础
In order to clone neurotrophic factor and trophic factor gene during nerve injury and regeneration, sciatic nerve tissue damaged by SD rats was used as material to extract mRNA and cDNA was reverse transcribed. A cDNA library was constructed with λgt11 as the carrier. The capacity of the library was determined to be greater than 1.5 × 106, the percentage of recombination was 96.7%, and the length of insert was between 0.5-8 kb. The titer after amplification reached 1.4 / 1012 pfu / ml. Using this library DNA as a template, 360 bp NGF and 616 bp CNTF reading frame fragments were amplified by PCR. CNTF clones were screened from the library using digoxigenin-labeled CNTF probes. DNA sequencing confirmed that the full sequence of NGF and CNTF cDNA. Based on the obtained 65kD chemically-induced factor antibody, the library was screened by ABC method and the positive clones were obtained. The inserted fragment was about 1.1kb. Genbank query confirmed as a new sequence. The establishment of this library lays the foundation for screening and isolating the gene of interest and further studying the mechanism of nerve regeneration, development and degeneration from the gene level