日本血吸虫黏蛋白样蛋白部分基因的扩增及测序

目的　体外扩增日本血吸虫中国大陆株黏蛋白样蛋白(SjMLP)抗原的部分基因序列。　方法　利用PCGENE软件查找SmMLP的抗原决定簇;特定寡核苷酸引物的设计与合成;Trizol抽提日本血吸虫成虫总RNA,逆转录聚合酶链反应(RTPCR)扩增目的基因,测序后与SmMLP进行同源性分析。　结果　RT PCR特异性扩增出SjMLP编码区基因序列,其片段大小为756bp,测序结果与SmMLP具有高度同源性。　结论　RTPCR扩增的SjMLP抗原编码区基因序列与预期相符合。

Amplification and Sequencing of the Gene Coding for Mucin-like Protein from Schistosoma japonicum

Objective To amplify and sequence the partial gene coding for mucin-like protein from Chinese isolates of Schistosoma japonicum (SjMLP). Methods The antigenic determinants of SmMLP were predicted by PCGENE software and specific oligonucleotide primers were designed and synthesized. Total RNA was isolated from adult worms of S.japonicum using Trizol reagent and the coding region gene of SjMLP was amplified by RT-PCR technique. Results The coding region of SjMLP gene was specifically amplified by RT-PCR and the size of amplified fragment was 756 base pairs. The DNA sequence analysis result indicated that the coding sequence of the MLP was highly homologous between S. mansoni and S.japonicum. Conclusion The amplified fragment is consistent to the predicted one, providing a basis for cloning and further study on DNA immunization.