穿梭载体(sbuttle vector)指在两种宿主生物体内复制的载体分子,因而可以运载目的基因(穿梭往返两种生物之间)。穿梭质粒含原核和真核生物 2 个复制子,以确保两类细胞中都能扩增。 科淼生物的各种产品发货前经经过严格的多重验证,收到质粒后请短暂离心,加入20ul ddH2O溶解质粒,取2ul转化至对应感受态中,挑取单克隆重新...
These two type-fragments were rep- licated three times and fused to the pAbAi vector, while CDS of PtoRD26.1 gene was cloned to the pGADT7 vector to create the AD-PtoRD26.1 vector. The AD and AD- PtoRD26.1 vectors with the RE2 motif or its mutant were then transformed into the Y1...
dram1 or dram1∆19N PCR products were ligated into a ZERO BLUNT TOPO vector (450245, ThermoFisher) and the insert was confirmed by Sanger sequencing (Base Clear, Netherlands). dram1 and dram1∆19N mRNA was generated using the SP6 mMessage mMachine kit (AM1340, Thermo Fisher) and ...
PtoRD26directly and positively regulatesSRMTexpression.AThe constructs of the effector and reporter in the dual-luciferase assay. The pCXSN empty vector contains accdBgene that is a negative selection marker to eliminate the self-ligation background after transformation.BPtoRD26.1enhanced fluorescence inten...
Bacterially expressed caspases with C-terminal His6 tags were purified on Talon affinity resins and then eluted with 250 mM imidazole in CHAPS buffer (20 mM HEPES, pH 7.0, 10 mM KCl, 0.1% CHAPS). Bacterially expressed WT DFNA5, DFNA5-D270E and GSDMD with N-terminal His6-T7 tags...
gene) must first be precloned into the expression vector. This prerequisite would mean an extra step when applying MEGAWHOP compared to PTO-QuickStep. However, the advantage is a higher number of transformants due to better annealing of the megaprimers (the complementary region is essentially the...