CAJ | 학술논문

目的构建日本血吸虫凝溶胶蛋白(Sjgelsolin)的原核表达载体,获得其表达产物并分析其在不同发育阶段的转录水平.方法根据序列AY814387设计引物,体外扩增Sjgelsolin的蛋白质编码序列,将其插入pET28a(+)质粒,转化至大肠埃希菌BL21(DE3),用IPTG诱导表达.提取日本血吸虫虫卵,尾蚴,7-d童虫,42-d雌、雄成虫的总RNA,用半定量RT-PCR分析Sj gelsolin在每个样本中的转录水平.结果成功构建了pET28a(+)-Sjgelsolin重组质粒,并获得高水平的表达.Sjgelsolin转录子在尾蚴和雌虫成虫中未被检测到,而在7-d童虫和42-d雄虫成虫中被检测到.结论 Sjgelsolin在体外获得表达,且其转录是受发育调控的.
목적 구건일본혈흡충응용효단백(Sjgelsolin)적원핵표체재체,획득기표체산물병분석기재불동발육계단적전록수평.방법 근거서렬AY814387설계인물,체외확증Sjgelsolin적단백질편마서렬,장기삽입pET28a(+)질립,전화지대장애희균BL21(DE3),용IPTG유도표체.제취일본혈흡충충란,미유,7-d동충,42-d자、웅성충적총RNA,용반정량RT-PCR분석Sj gelsolin재매개양본중적전록수평.결과 성공구건료pET28a(+)-Sjgelsolin중조질립,병획득고수평적표체.Sjgelsolin전록자재미유화자충성충중미피검측도,이재7-d동충화42-d웅충성충중피검측도.결론 Sjgelsolin재체외획득표체,차기전록시수발육조공적.
Objective To construct prokaryotic expressional vector of gene gelsolin from Schistosoma japorticum(Sjgelsolin),obtain its expressional product and analyze its transcriptional levels in different life stages.Methods Primers were designed based on the sequence of AY814387,protein-coding sequence of sjgelsolin wag amplified in vitro,inserted into pET28a(+)plasmid,and transformed into E.coli BL21(DE3) cells,and then expressed under the induction of IPTG.Total RNAs were purified from egg,cercariae,7-day old larval worms,42-day-old adult female and male worms of Schiswsoma japonicum,and the transcriptional levels of gelsolin in each sample were analyzed by semi-quantitative RT-PCR.Results Recombinant plagmid pET28a(+)-Sjgelsolin wag successfully constructed and highly expressed.Sjgelsolin transcripts were not detected in cereariae or adult female worul8.but were observed in egg.7-day-old larval and adult male worms.Conclusion Sjgelsolin Wag succesfully expressed in vitro,and its transcription was developmentally regulated.