具重复Kringle结构分子的缺失体和嵌合体的构建方法

Construction strategy of serious deletants and chimeras of molecules with multiple Kringles

确定肝细胞生长因子(HGF)和巨噬细胞刺激蛋白(MSP)各4个Kringle结构的共有序列并设计公用引物,此公用引物配合HGF和MSP上、下游引物,利用DNA聚合酶链式反应(PCR)技术,可在一次PCR反应中扩增出多条片段,以不同的组合方式将这些片段两两连接,得到了缺失不同Kringle的缺失体和两种分子间不同区域的缺失型嵌合体,从而为HGF和MSP结构与功能的研究奠定了基础。此方法可普遍适用于含有两个或两个以上Kringle分子的缺失体与嵌合体的构建。克隆过程中对存在引物酶切位点的cDNA片段的克隆方法的建立,为同类问题的解决提供了可靠的方法。

According to the sequence conservation of four Kringles of HGF and MSP, the universal primers should be designed. With these primers for PCR, various segments with different length may be obtained at one PCR and used to construct serious deletants and chimeras of HGF and MSP. Then the functions of these deletants and chimeras should be observed. As a result, the function determines should be figured out.