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从原代培养人许旺细胞申提取总RNA，用RT－PCR方法获取了编码带向导肽和成熟的NT-3两个cDNA片段，经DNA序列测定表明其顺序与文献报道一致。将这两个cDNA片段克隆到大肠-酵母穿梭质粒pVT102U／α上。用酵母整体细胞法将重组质粒转化酵母宿主细胞，经筛选后，获得分泌性表达。这两种cDNA片段的表达产物经SDS-PAGE电泳鉴定，大小都在15ku左右，说明带前导肽的NT-3在酵母细胞中获得了正确的加工。将这两种部分纯化的NT－3样品经鼠胚背根神经节生物活性分析，表明具有明显的促进细胞存活和突起生长的作用。人神经营子-3％在酿酒酵母系统获得有生物活性的表达。
Total RNA was extracted from primary cultured human Schwann cells. Two cDNA fragments coding for the guide peptide and mature NT-3 were obtained by RT-PCR. The DNA sequence showed that the order was consistent with that reported in the literature. Both cDNA fragments were cloned into the large intestine-yeast shuttle plasmid pVT102U / α. Recombinant plasmids were transformed into yeast host cells by yeast whole cell method and screened to obtain secretory expression. The expression products of these two cDNA fragments were identified by SDS-PAGE electrophoresis and the size was around 15ku, indicating that the leader peptide NT-3 was correctly processed in yeast cells. Analysis of the biological activity of these two partially purified NT-3 samples from the dorsal root ganglion showed that the NT-3 partially purified NT-3 had obvious effects of promoting cell survival and neurite outgrowth. Human neurotrophic factor -3% in Saccharomyces cerevisiae system to obtain the biological activity of expression.