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为了提高人睫状神经营养因子（ＣＮＴＦ）的生物学活性，用ＰＣＲ方法获取Ｎ端缺失１４个氨基酸的ＣＮＴＦ基因片段，经酶切鉴定、核酸测序证实突变体的核苷酸序列，将其重组至表达质粒ｐＢＶ２２０，构建了ＣＮＴＦ突变体表达载体ｐＢＶ－ＣＮＴＦΔ．用ＳＤＳ－ＰＡＧＥ测定其表达水平，鸡胚背根节无血清培养法检测表达蛋白的生物学活性．结果表明，ｐＢＶ－ＣＮＴＦΔ能表达生物学活性高于天然ＣＮＴＦ的约２６ｋＤ蛋白质，表达水平达３０％．为今后通过基因工程方法获得ＣＮＴＦ突变体，从而制备高效的ＣＮＴＦ制剂创造了条件．
In order to improve the biological activity of human ciliary neurotrophic factor (CNTF), a 14-amino acid deletion fragment of CNTF was obtained by PCR. The nucleotide sequence of the mutated CNTF was confirmed by restriction enzyme analysis. To express plasmid pBV220, a CNTF mutant expression vector pBV-CNTFΔ was constructed. The expression level of the expressed protein was detected by SDS-PAGE. The biological activity of the expressed protein was detected by serum-free culture of chicken embryo dorsal root ganglion. The results showed that pBV-CNTFΔ could express about 26kD protein with biological activity higher than that of native CNTF, and the expression level was up to 30%. For the future through genetic engineering methods to obtain CNTF mutants, thus preparing highly efficient preparation of CNTF conditions.