MethodsNucleotides within glycosylation motif were changed to a human mutant CD9 gene(from K-H to K-K) by overlap extension PCR. The mutant CD9 gene was inserted into a eukaryotic-expressing vector pALTER to form a recombinant plasmid.
MethodsNucleotides within glycosylation motif were changed to a human mutant CD9 gene(from K-H to K-K) by overlap extension PCR. The mutant CD9 gene was inserted into a eukaryotic-expressing vector pALTER to form a recombinant plasmid.
方法以正常的CD9基因全长cDNA序列为模板,应用重叠延伸PCR法产生突变使CD9糖基化基序K-H突变为K-K,产生突变CD9基因,将其克隆于pALTER质粒中,构建出重组真核表达载体。