日本血吸虫真核表达质粒pcDNA3/HGPRT的构建及序列测定

目的应用基因工程技术,对日本血吸虫次黄嘌呤-鸟嘌呤磷酸核糖转移酶(hypoxanthine-guaninephosphoribosyltransferase,HGPRT)进行亚克隆,试图构建真核重组DNA质粒pcDNA3-HGPRT。方法通过筛选日本血吸虫成虫cDNA文库,克隆HGPRT基因片段,然后将目的片段亚克隆入真核表达载体pcDNA3。结果成功构建了真核重组DNA质粒pcDNA3-HGPRT,为进一步研究HGPRT的功能奠定了基础。

Construction and sequencing of pcDNA3-HGPRT of Schistosomiasis japonicum

Aim In order to construct the recombination plasmid by pcDNA3 vector with hypoxanthine guanine phosphoribosyl transferase (HGPRT) of Schistosomiasis japonicum.Methods HGPRT is one proteins selected from the SIEA components as target with high homogeneity and immunogenicity identified through MALDI TOF MS analysis. Its structures and functions including physical characteristics,average molecular weight,HPLC features,hydrophobicity,average flexibility,α helix,beta turn and beta sheet etc. will report on the next papers. To search for the genes coding HGPRT,the S.japonicum adult worm cDNA library was screened. Results we identified the result of pcDNA3 HGPRT from PCR amplification with its gene from S.japonicum adult worm cDNA library after repeated screenings and sequencing.Conclusion In summary,the results provide some important experimental bases for advanced exploration of HGPRT as a molecular vaccine against Schistosomiasis japonicum.