利用套叠PCR技术进行基因突变和拼接

利用套叠PCR技术（又称重叠区扩增基因拼接法）对hGM-CSF基因内第28位氨基酸处的糖基化位点进行突变和进行人促性腺激素基因，腺苷酸激酶短肽与胰岛素样生长因子－基因三者之间的拼接，结果表明采用该技术能在体外实行有效的基因重组和定点突变，其成功率为100％，这一技术不需要内切酶消化和连接酶处理，技术操作员简单易行，在基因拼接，基因内部突变方面具有良好的应用价值。

Recombination and Mutation of some Genes with Gene Splicing by Overlap Extension

The glycosylation site (at the 28th amino acid residue) of human granulocyte-macrophage colony stimulating factor (hGM-CSF) gene was mutated by gene SOEing.Follicle stimulating hormone gene (FSH)，short peptide of adenine nucleotide kinase and insulin-like gowth factor-I gene (IGF-I) were recombined together by gene splicing by overlap extension (gene SOEing). Results showed that gene SOEing was a successful specific PCR technique for gene recombination and gene mutation in vitro. The technique didnt need restrict enzymes and ligases，which could be developed as a simple and useful technique in the studies of gene splicing and gene mutation.