华支睾吸虫HMGB1同源分子的识别及其进化研究

Identification and evolution of HMGB1 homologous molecules of Clonorchis sinensis

目的识别华支睾吸虫HMGB1同源分子的CDS及其蛋白序列,并分析其进化特征。方法分别用基于麝猫后睾吸虫HMGB1预测序列的5'和3'RACE以及基于华支睾吸虫HMGB1基因组预测序列的PCR法扩增华支睾吸虫成虫来源的cDNA,对扩增产物进行克隆测序,并进行序列的同源性比较和拼接,分析其ORFs,据此对华支睾吸虫成虫来源的cDNA进行PCR扩增验证其转录;构建进化树,对识别出的HMGB1同源分子进行进化分析。结果识别出两个华支睾吸虫HMGB1同源蛋白CDS,大小分别为381和477 bp,二者均在华支睾吸虫成虫转录,产物均一;推测其编码蛋白分别为127和159个氨基酸,其分子序列与血吸虫同源性较高,而与人及其他脊椎动物同源性低。结论成功识别出华支睾吸虫HMGB1同源蛋白CDS序列,为研究其与肝胆管癌发生的关联性和机制奠定了基础。

Objective To identify the CDS and its protein sequences of the HMGB1 homologous molecule from C.sinensis,and to analyze its evolutionary features.Methods The C.sinensis adult derived cDNA was amplified by 5'and 3'race based on the predicted sequence of O.viverrine HMGB1 and by PCR based on the predicted sequence of C.sinensis HMGB1 genome,respectively.The amplified products were clonally sequenced,and the sequences were compared for homology and splicing,and their ORFs were analyzed,according to which the transcription was verified by PCR amplification of cDNA derived from the adult worms.An evolutionary tree was constructed,and the identified HMGB1 homologous molecules were subjected to evolutionary analysis.Results Two C.sinensis HMGB1 homologous protein CDS,381 and 477 bp in size,were recognized,both of which were transcribed in C.sinensis adults with uniform products.Its encoded proteins are presumed to be 127 and 159 amino acids,respectively,and its molecular sequence shares high homology with schistosomes but low homology with humans and other vertebrates.Conclusion The CDS sequences of the HMGB1 homologous protein from C.sinensis were successfully identified,which laid the foundation for investigating their association and mechanism with hepatobiliary carcinogenesis.