杀菌通透性增加蛋白基因的克隆及在CHO细胞中的表达

GENE CLONING AND EXPRESSION OF HUMAN BACTERICIDAL/ PERMEABILITY-INCREASING PROTEIN

从一名健康中国人外周血白细胞中克隆出杀菌 通透性增加蛋白 (BPI)基因 ,全长1 45 2bp,编码 2 7个氨基酸的信号肽和 45 6个氨基酸成熟蛋白。序列比较发现 ,此序列与国外发表的序列有 6个核苷酸的差异 ,编码蛋白有 4个氨基酸的差异。构建真核表达质粒 ,在CHO细胞中实现BPI基因的稳定表达。对重组蛋白初步纯化后进行杀菌实验 ,结果表明重组蛋白具有与天然蛋白同样的生物活性。

The gene of human bactericidal/permeability-increasing protein (BPI) was cloned from peripheral blood lymphocytes of a normal Chinese individual.The result of sequencing showed the gene is 1452bp encoding a 27-residue signal peptide and a 456-residue matured protein, and it has six nucleotide variations compared with the sequence reported which results in 4 different amino acids.In order to get recombinant BPI, the gene was cloned into an expressing plasmid and expressed in CHO cells. The recombinant protein was purified using cation-exchange chromatography and its bioactivity was proved with bactericidal assays.