人细菌透性增加蛋白cDNA的克隆及序列分析

cDNA Cloning and Sequence Analysis of Human Bactericidal/Permeability-increasing Protein

摘要:

从新疆维吾尔族健康人造血干细胞中提取总RNA,用反转录(reverse transcription,RT)和降落PCR (touchdown PCR,TD-PCR)相结合的方法扩增人细菌透性增加蛋白(human bactericidal/permeability-increasing protein,hBPI)的cDNA,将其克隆到pEGFP-N1载体上并进行DNA序列测定。结果表明,所克隆的hBPIcDNA全长为1,464个碱基,其序列与GenBank中另外4个序列进行了比对,有两个碱基与其它4个序列不同:其中第576位碱基其它序列为G,该位置碱基是C;第676位碱基其它序列为A,该位置碱基是G。其中第676位碱基的变化导致第185位氨基酸由Lys改变为Glu。

关键词:人细菌透性增加蛋白; cDNA; TD-PCR; 基因克隆; DNA序列分析

Abstract:

Human bactericidal/permeability-increasing protein(hBPI)cDNA was amplified by reverse transcription(RT)and touchdown PCR(TD-PCR)from blood stem cells collected from healthy human of Uygur nationality in Xinjiang Uygur Autonomous Region of China, and then was subcloned into pEGFP-N1 vector,hBPI cDNA sequence consists of 1,464bp.Comparison with other 4 hBPI cDNA sequences registered in GenBank identified 99% homology in DNA sequence.However,there were two base substitutions(nucleotide 576G→C,nu- clotide 676A→G),one of which resulted in an amino acid (residue 185 Lys→Glu).

Key words:Human bactericidal/permeability-increasing protein; cDNA; TD-PCR; Gene cloning; DNA sequence analysis