布氏锥虫C型凝集素类似蛋白的鉴定

Identification of C-type lectin-like proteins of Trypanosoma brucei

C型凝集素在机体免疫系统中具有重要的作用。为了解布氏锥虫(T.brucei)基因组编码的C型凝集素类似蛋白(CLLPs),本研究在GeneDB数据库中通过软件鉴定了13种T.brucei基因组编码的CLLPs家族基因序列,并分析它们之间的遗传进化关系。同时,采用RT-PCR方法检测了其中3种CLLPs基因在血液阶段(BSF)T.brucei虫体内的转录。利用PCR扩增并在大肠杆菌中对所选3种基因进行表达,以表达的重组蛋白免疫小鼠制备多克隆抗体。实验结果表明,本研究选择的3种CLLPs基因在BSF T.brucei虫体内均能够进行转录;而通过间接免疫荧光检测表明所选的3种CLLPs基因中,只有一种CLLP(Tb5290)在T.brucei的BSF阶段和昆虫阶段(PCF)表达。本研究首次在T.brucei中鉴定了C型凝集素家族的新成员,为进一步了解CLLPs在T.brucei中的生物学功能的研究奠定了基础。

C-type lectins play important roles in the immune system. In the present study, thirteen different sequences encoding the C-type lectin-like proteins （CLLPs） of Trypanosoma brucei were identified in the GeneDB by bioinformatic softwares. In addition, the phelogenetic relationships of the CLLP sequences were analyzed and the transcription of 3 CLLPs in blood stream form （BSF） of T.brucei were detected by RT-PCR. Three of the CLP genes were amplified from the T.brucei genomic DNA and expressed in E.coli to prepare the polyclonal antibodies. The results showed that the 13 CLLP sequences were classified into 3 groups. The transcription of the 3 CLLPs were detected in BSF of T. brucei. However, indirect fluorescence assay indicated that, only one of the three genes expressed in BSF and procyclic form parasites. In conclusion, new members of C-type lectins were identified in this study, which would facilitate for the further research of the biological functions of proteins in T.brucei.