日本血吸虫大陆株泛蛋白缀合酶E(SjUCEE)基因的A-T克隆

目的　日本血吸虫中国大陆株泛蛋白缀合酶 E (ubiquitin- conjugating enzyme E ,Sj UCEE)的基因克隆和鉴定。方法　特定寡核苷酸引物 PCR扩增目的基因 ;应用分子克隆常规操作方法将扩增产物克隆至载体 pu Cm - T中。结果　 PCR特异性扩增出 Sj UCEE编码区基因序列 ,其片段大小为 75 7bp;经酶切、PCR及测序鉴定表明所构建的质粒 pu Cm - T/Sj UCEE中含有所扩增的基因序列。结论　 PCR扩增的 Sj U CEE抗原编码区基因序列与预期长度相符合 ,成功构建了含目的基因的原核质粒 pu Cm- T/Sj U CEE

A-T CLONING OF GENE CODING UBIQUITIN-CONJUGATING ENZYME E FROM CHINESE SCHISTOSOMA JAPONICUM

Objective To clone and identify ubiguitin-conjugating enzyme E gene of Schistosoma japonicum Chinese strain (SjUCEE).Methods The specific oligonucleotide primers were designed and synthysized. The extraction,coding region gene of SjUCEE was amplified by PCR technique and the gene from PCR was cloned into cloning vector puCm-T. The resulting construct was determined by PCR, restriction and sequence analysis methods. Results The coding region of SjUCEE gene was specifically amplified by PCR and the size of amplified fragment was 757 base pairs.The DNA sequence analysis indicated that the puCm-T/SjUCEE plasmid included the coding sequence of the SjUCEE. Conclusion The amplified gene is identical with the predicted one and the puCm-T/SjUCEE plasmid is successfully constructed.