简介:VariableChargeX/Y(VCX/Y)isahumantestis-specificgenefamilythatlocalizedonXandYchromo-somes.Inthisstudy,VCYproteinwasexpressedinE.coliintheformofglutathione-S-transferase(GST)fusionprotein.Withthepurifiedfusionproteinasantigen,theanti-GST-VCYantibodywasgeneratedandthelocalizationofVCYproteininhumantestiswasdeterminedbyimmunohistochemistry.Inthetestisseminiferousepithelium,VCYproteinswerehighlyexpressedinnucleiofgermcells.Usingpropidiumio-didestainingandgreenfluorescentprotein(GFP)tagtechnologies,VCYandVCX-8rproteinsweremainlylocalizedinthenucleoliofCOS7cells.Inaddition,thecolocalizationforVCYandVCX-8rinCOS7cellswasalsoobserved.WithVCYcDNAasbait,acDNAfragmentofacidicribosomalproteinPOwasobtainedusingyeasttwo-hybridsystem.AlltheinformationaboveindicatesthatVCX/Yproteinfamilymightbeinvolvedintheregulationofribosomeassemblyduringspermatogenesis.
简介:【摘要】在深圳卷烟厂,YJ112卷烟机已作为卷烟卷制环节的主力机型;该机型主要依靠平准器对烟支重量进行控制调节和紧固烟支端部烟丝。通过对YJ112卷烟机平准器主传动法兰部位的改进,采用主传动法兰表面喷涂陶瓷和主传动法兰内孔加装防油垫片等措施,有效减少了平准器漏油的次数;通过安装双重法兰和回油管,进一步降低了平准器漏油风险,加装回油管使得漏油更容易观察;通过实践证明,以上的设计改进是可行的。
简介:摘要目的构建细粒棘球蚴pET30a-EgG1Y162-2原核表达重组质粒,诱导表达EgG1Y162-2重组蛋白,为研制细粒棘球蚴疫苗提供研究基础。方法以细粒棘球蚴cDNA为模板,利用PCR法合成EgG1Y162-2目的基因,经限制性内切酶EcoRⅠ和HindⅢ双酶切后连接到原核表达载体pET30a中,构建pET30a-EgG1Y162-2重组质粒;将重组质粒转化至大肠埃希菌BL21(DE3)感受态细胞中,经异丙基硫代半乳糖苷(IPTG)诱导表达大量蛋白,采用亲和层析方法纯化重组蛋白;经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分析蛋白纯化水平,蛋白免疫印迹(Western blot)法鉴定表达产物。结果成功构建pET30a-EgG1Y162-2重组质粒,诱导表达后菌体上清液和洗脱液均在相对分子质量约15 × 103处出现EgG1Y162-2蛋白目的条带,且200 mmol/L咪唑洗脱的蛋白抗原成分较纯。Western blot结果显示,纯化后带有His标签的EgG1Y162-2重组蛋白能被抗His单克隆抗体识别。结论成功构建了细粒棘球蚴pET30a-EgG1Y162-2原核表达重组质粒,诱导表达了EgG1Y162-2重组蛋白,为进一步开展抗细粒棘球蚴疫苗的研究奠定基础。