论文部分内容阅读
目的 为了进一步研究日本血吸虫 (中国大陆株 ) Sj16的免疫调节功能 ,丰富有关血吸虫免疫逃避知识。 方法 根据曼氏血吸虫 Sm16基因已知序列设计合成一对引物 ,用 PCR技术从日本血吸虫 (中国大陆株 )成虫 c DNA文库中扩增 Sj16基因 ;将 Sj16基因定向克隆入 pc DNA3,转化感受态 DH5α菌 ;用酶切、PCR扩增鉴定筛选得到的重组阳性克隆。以重组 pc DNA3- Sj16质粒为模板 ,用双脱氧链末端终止法测定 Sj16基因序列 ,应用软件辅助分析 Sj16序列及进行 Sj16与曼氏血吸虫的 Sm16同源性比较。 结果 从日本血吸虫 (中国大陆株 )成虫 c DNA文库中获取 Sj16基因 ,重组质粒中含有 Sj16基因 ,成功构建日本血吸虫 (中国大陆株 )Sj16基因真核表达重组质粒 pc DNA3- Sj16 ;Sj16基因开放读码框有 35 4碱基 ,编码 117氨基酸 ,N端 18个氨基酸可能为信号肽序列 ;Sj16与 Sm16有高度同源性 ,只存在一个氨基酸的差异。 结论 成功克隆了 Sj16基因的真核表达载体 ,并测定、分析了 Sj16基因序列 ,为深入研究 Sj16的免疫调节功能 ,丰富有关血吸虫免疫逃避知识打下基础。
Objective To further study the immunomodulatory function of Sj16 from Schistosoma japonicum (Chinese mainland strain) and enrich the immune evasion knowledge of schistosomiasis. Methods A pair of primers was designed and synthesized based on the known sequence of Sm16 gene of Schistosoma mansoni. Sj16 gene was amplified from adult c DNA library of Schistosoma japonicum (Chinese strain) by PCR. Sj16 gene was cloned into pc DNA3 and transformed into competent DH5α Bacteria; digestion, PCR amplification identified positive recombinant clones. The Sj16 gene sequence was determined by the dideoxy chain termination method using the recombinant pcDNA3-Sj16 plasmid as a template. Sj16 sequence was analyzed by software and the homologies of Sj16 to Sm16 of Schistosoma mansoni were compared. Results The Sj16 gene was obtained from the adult c DNA library of Schistosoma japonicum (Chinese mainland strain), and the recombinant plasmid contained Sj16 gene. The recombinant plasmid pcDNA3-Sj16 was successfully constructed. The Sj16 gene was successfully readjusted for Sj16 gene of Schistosoma japonicum (Chinese mainland strain) There are 35 4 bases in the code frame, encoding 117 amino acids. The 18 amino acids at the N terminal may be signal peptide sequences. Sj16 and Sm16 have high homology with only one amino acid difference. Conclusion The eukaryotic expression vector of Sj16 gene was successfully cloned and sequenced. The sequence of Sj16 gene was determined and analyzed. The purpose of this study was to further investigate the immunomodulatory function of Sj16 and enrich the knowledge of immune evasion of schistosomiasis.