Balb/c小鼠甘露聚糖结合凝集素-C基因的克隆与鉴定

Molecular cloning and characterization of mannan binding lectin C gene from Balb/c mice

目的获得小鼠甘露聚糖结合凝集素-C(MBL-C)基因的全长编码区cDNA。方法利用RT-PCR方法,从Balb/c小鼠的肝细胞中分离出MBL-C基因cDNA片段,将其克隆入pUC-T载体并测序,对基因结构及其与人MBL基因的同源性进行比较分析。结果扩增得到的Balb/c小鼠MBL-C基因cDNA全长735 bp,编码245个氨基酸残基,包含了完整的编码区基因序列。经核苷酸序列比较分析发现,扩增得到的Balb/c小鼠MBL-C基因与Genbank中发表的序列具有100%的同源性,与人的MBL基因的同源性为71.4%。结论成功地克隆到完整的Balb/c小鼠MBL-C编码区基因,为进一步在整体水平研究MBL分子的天然免疫功能提供了必要的条件。

Objective To obtain full-length cDNA encoding mouse mannan-binding lectin C(MBL-C) polypeptide. Methods The cDNA encoding mouse MBL-C was isolated from Balb/c mouse liver cells by reverse transcriptase (RT)-PCR and insert-ed into pUC-T vector, and the encoding region structure of the DNA was analyzed to understand its evolutionary relationship with single human MBL homologues. Results The amplified mouse MBL-C cDNA, which was 735 bp in length, encoded 245 amino acid residues, and its structural analysis showed 100% homology with published mouse MBL-C gene sequence and 71.4 % homology with MBL gene of Chinese human. Conclusion The gene encoding Balb/c mouse MBL-C has been success-fully isolated, which may facilitate further study of the innate immune functions of MBL molecule in vivo.