To this end, we replaced all cytidine residues of a 27 nt HIV-1 TAR RNA mimic by the 6-C-modified counterparts and added a TEMPO moiety at the 5′-terminus yielding construct 6.
Fresh fd-RP vector is the shorter vector prepared by first digesting the fd-RP DNA with BamHI and ligating the resultant vector fragment, yielding a construct with a truncated SfiI fragment [18].
The complete sequence of Xenopus laevis Eg5 (corresponding to the complete reading frame from amino acid 1 to 1067, gift of Claire Walczak) was amplified by PCR and inserted into pFASTBacHTa (Invitrogen) using the SalI and NotI sites yielding a construct with a N-terminal hexahistidine tag.
Cytochrome c (cytc) and its adduct with cardiolipin (CL) were immobilized on a hydrophobic SAM-coated electrode surface yielding a construct which mimics the environment experienced by the complex at the inner mitochondrial membrane where it plays a role in cell apoptosis.
Cysteine residues were engineered into MBP · CaM (MBP-T34C,T110C-CaM) and labeled with donor and acceptor fluorescent probes yielding a construct (MBP · CaM-DA) which can be used for single-molecule single-pair fluorescence resonance energy transfer (spFRET) experiments.
The KcsA sequence was cloned into a pET-30 Ek/LIC expression vector (Merck-Novagen, Darmstadt, Germany), yielding a DNA construct encoding a fusion protein comprising a hexahistidine tag, a 44-residue spacer, and KcsA residues 1 160 (ref (40)).
Some selected SPs were also ligated into the 6.9 kb NdeI- SalI fragment of pUsp45-Amy [ 19], yielding constructs for secretion of AmyA.
The SP-containing fragment was excised from the resulting plasmid by NdeI- SalI restriction digesting and ligated into the 6.1 kb NdeI- SalI fragment of pUsp45-Nuc[ 19], yielding constructs for secretion of NucA.
Two independent single-end cDNA libraries of sample A were constructed and sequenced, yielding datasets 'A' that contained ~29.7 million reads and 'A-rep' that was sequenced at 3-fold lower depth and contained ~9 million reads.
In sum, cognitive plasticity seems to be a relatively new construct yielding useful information about normal and impaired mental functioning being influenced by age, level of pathology, and education.
Briefly, PCR amplified aquaporin cDNA fragments were co-transformed into YSH1770 strain with BamHI-, HindIII- and SalI-digested pUG35 for synthetic cDNA-derived hAQP10 and its variants, or SpeI- and ClaI-digested pUG35 for genomic cDNA-derived hAQP3 and GFP PCR products yielding hAQP3GFP construct.
