简介：TherootofPanaxginsengplantundergoesaspecificdevelopmentalprocesstobecomeabiosynthesisandaccumulationtissueforginsenosides.Toidentifyandanalyzegenesinvolvedinthebiosynthesisofginsenoside,weconstructedandcharacterizedafull-lengthcDNAlibraryfor6-year-oldNorthAmericanginseng.ThetiterofprimarycDNAlibraryis1.2×106pfu/mL,thetiterofamplifiedlibraryis2.6×1010pfu/mLandtherateofrecombinantisabove86%.Theinsertsizerangesfrom0.3to2.0kb.Sequencingresultsshowthat18of58genesarehighhomologoustothegenes(GBR5,GBR3andGBR1)knowninGenBank,whichareinvolvedinbiosynthesisofginsenosideinNorthAmericanginsengplant;16of58genesarenovelgenes.Thefull-lengthcDNAlibraryofNorthAmericanginsengroottissuesisessentialforthecloningofgenesknownanditisalsoaninitialkeyforthescreeningandcloningofnewgenes.

简介：用PCR方法扩增了人的BAF53全长cDNA序列,用酶切后的ΔBAF53cDNA片段构建了表达质粒.转化大肠杆菌后,经IPTG诱导表达,再经包涵体裂解、纯化,得到hBAF53抗原多肽.从免疫动物身上获得实验动物的抗血清.通过斑点印迹方法测得hBAF53抗血清的特异性和效价,又用已提取的HeLa细胞核蛋白(含有天然BAF53蛋白)进行免疫印迹分析,证明天然BAF53蛋白是该抗血清的抗原,说明获得的抗血清具有比较高的特异性,可用于染色体改构复合物各亚基间相互作用分析及调控基因转录机理方面的研究.