狼毒大戟为大戟科多年生草本植物.本研究以狼毒大戟(Euphorbia fischoriana)为试验材料,构建狼毒大戟cDNA文库及EST测序.研究结果表明,我们成功地构建了狼毒大戟的cDNA文库,并获得了2099个EST的序列.进一步对这些ESTs数据进行了Unigene的拼接、ORF预测和相应的功能注释及分类,发现了一些可能参与Prostratin生物合成的候选结构基因Transketolase,NADPH-cytochrome P450 reductase,Cytochrome P450,S-adenosylmethionine synthetase,acetyltransferas,l-deoxy-D-xylulose-5-phosphate synthase (DXP)和候选调节基因AP2类ERF转录因子(APE-like ethylene-responsive transcription factor,AP2-like ERF),此研究为今后开展抗HIV次生代谢物Prostratin的代谢工程奠定了基础.
原发性胆汁性肝硬化(primary biliary cirrhosis,PBC)是一种慢性渐进性胆汁淤积性肝脏疾病,以小胆管破坏为主要组织学特征的非化脓性炎症,并以血清中出现特征性自身抗体为主要标志.自身抗体对该病发病机制、诊断、治疗及预后均有重要意义.我们就PBC自身抗体谱研究进展作简要综述.
AIM: To clone and construct the plasmid containing human autoantigene Sp100 gene, and then to identify the immunoreactivity of the purified recombinant protein. METHODS: The Sp100 gene was amplified from human liver cDNA library, and then was cloned into PEGH vector to induce the Sp100 expression. The obtained products were identified and analyzed by sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE) and Western blot. RESULTS: The sequence of Sp100 autoantigene gene was confirmed to be the same as the sequence reported in GenBank. The fusion proteins were found at 53-, 55-, 52-, 37-, 42-, and 47-ku strip on SDS-PAGE gel. Western blot analysis showed that the fusion protein with 55, 52, 42 and 47 ku had the same immunoreactivity as human Sp100 autoantigene. CONCLUSION: Human plasmid containing SplOO gene is successfully cloned and expressed in yeast Y258.
