PCR技术对人IL-3 cDNA进行定点突变的研究

PCR Site-Directed Mutagenesis of Human IL-3 cDNA in vitro

本文利用PCR技术对人IL-3cDNA体外进行定点突变,将人IL-3cDNA第3位Met,第116位Lys密码子突变为Val密码子GTT。PCR扩增片段核苷酸序列与引物设计相应的cDNA突变体序列完全一致。结果证实此方法比经典寡聚核苷酸方法简单、迅速、成本低、效率高,也为基因的修饰,蛋白质工程研究提供了简便、稳定的方法。

The human IL-3 cDNA mutants were obtained by PCR in vitro. Nucleotide sequence? of their PCR products were determined by DNA sequence analysis. The codon Met3 and Lys116 of human IL-3 cDNA were substituted by codon (GTT) of Val. PCR site-directed mutagenesis method is simpler and faster than classical mutagenesis method. We describe the method of PCR site-directed mutant and it is efficient, cost effective, high-fidelity. The method is used modification of gene, protein to study structure/ function relation.