简介:建立了商业化种植的NeWLeaf-YTM转基因马铃薯的筛选检测和鉴定的PCR方法.该方法根据马铃薯自身patatin基因作为内源特异参照基因扩增216bp片段,检查模板DNA提取的质量,避免了假阴性结果;同时扩增FMV35S启动子基因225bp片段和PVY-cp基因161bp片段.PCR反应循环参数是94℃2min;94℃40s,55℃60s,72℃60s,35次循环;之后72℃延伸5min.本实验中的F-primer(PVY01-5′)/R-primer(PVY01-3′)引物是针对商业化种植的NewLeaf-YTM转基因马铃薯PVY-cp抗病毒基因而设计的,具有很强的特异性,可以达到对NeWLeaf-YTM转基因马铃薯鉴定的目的.
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.