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...
(a) Immunoblots of cell lysates from stable 293T-DFNA5-EGFP and 293T-DFNA5-D270E-EGFP cells transfected with vector control (Ctrl) or constructs for Bax, constitutively active T7-tagged caspase-3 (casp-3) or inactive caspase-3 (casp-3-C285A) for 24 h as indicated. The blots were...
during Mm inf Fig. 1: Dram1 deficiency leads to increased susceptibility to Mm infection. aSchematic representation of the zebrafishdram1/Dram1(ENSDARG00000045561/ENSDARP0 0000066996.3) genetic and protein domain architecture and CRISPR/Cas9 target site. Dram1 (240 amino acids) contains six transme...
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...
The coding sequences ofPtoCDKswere cloned into thepGADT7vector. The coding sequence ofPtoCYCD3;3was cloned into thepGBKT7vector.Yeast strainAH109 was co-transfected withpGBKT7andpGADT7constructs carrying the corresponding genes in accordance with the yeast transformation kit (SK2400-200, Coolab...
PCR products were gel-purified and cloned using a pGEM-T vector system and sequenced using an ABI 3100 automated sequencer (Applied Biosystems, Foster City, CA, USA). 4.3. Identification and Structural Characterisation of Baltikinin from the Skin Secretion Lyophilized skin secretion (5 mg) was ...