This construct was amplified by PCR from the above version, with the C-terminal primer incorporating a further 4-Histidines. The resulting amplicon was then inserted into BamHI-XhoI cleaved pGEX-6P-1 by In-Fusion cloning. The PLpro expression construct was similarly optimised and synthesised and ...
The PCR product was cloned into NdeI/HindIII-cut pIJ10257 (Hong et al., 2005) to generate plasmid pIJ10902. This construction results in whiG being placed under the control of the strong constitutive ermE∗ promoter. For overexpression of both WhiH and WhiI together, the coding regions of...
The genomic DNA sequence was used to design PCR primers to amplify over the exons and intron/exon boundaries of the full length of the OATP gene and some regions upstream of exon 1 and downstream of the final exon. Twenty-nine individual Caucasian genomic DNA samples were used as templates ...
To generate the GFP-LacI-PRMT6 vector, the PCR product of the human PRMT6 gene was cloned into the GFP-LacI vector (pKG215, Addgene, Watertown, MA). Rabbit antibodies against GFP were generated previously64. For Western blotting, antibodies against the following were used (clone name, ...
Some Kidins220 mRNA were detected by PCR, however, this did not yield any protein.1 Western blot analysis showed complete Kidins220 loss in knockout brains (Figure 1E),1 whereas expression of other Kidins220 inter- actors, such as TrkB and TrkC,15 was unaffected (Figure 1E). Antibodies...
Download: Download full-size image FIGURE 5. HTRA1 transcription in human iPSCs derived from wet AMD patients. A, genotyping of non-insertion/deletion versus insertion/deletion versions of the human HTRA1 gene transcription regulatory region. B, detection of human iPSC marker genes by RT-PCR. ...
After PCR, the resulting products were subjected to electrophoresis by using 1.5% agarose gel electrophoresis and EB staining, then an ARMS-specific primer that can effectively distinguish the D. brymerianum among Dendrobium species was chosen based on the results of the PCR amplification. ...