Plasmid, sequencing vectors

Materials pJK502 is a T7-overexpression vector that encodes for wild-type E.coli RNase HI. It was made by site-directed mutagenesis of pSMlOl (24), reverting three alanines (residues 13, 63 and 133) to cysteines, and then subcloning the resulting gene into a pETlla overexpression vector (details of plasmid sequence available upon request). Overexpression and purification of E.coli RNase HI were performed essentially as described in (24) for RNase H. RNase H is... 

The construction of the recombinant adenovirus expressing human FGF-4 required three components the FGF-4 transgene a plasmid shuttle vector to carry the transgene as well as 5 Ad5 sequences and a second plasmid carrying the bulk of the Ad5 genome. 

Most proteins used for therapeutic purposes In humans or animals are secreted glycoproteins stabilized by disulfide bonds. When researchers first tried to synthesize such proteins using plasmid expression vectors in bacterial cells, the results were disappointing. In most cases, the proteins were not secreted (even when a bacterial signal sequence replaced the normal one) instead, they accumulated in the cytosol and often in a denatured state, in part owing to the lack of disulfide bonds. After It became clear that disulfide-bond formation occurs spontaneously only in the ER lumen, biotechnologists eventually developed expression vectors that can be used In animal cells (Chapter... 

XLl-Blue A(mcrA)183 A(mcrBC-hsdMRS-tnrr)173. A good general host for plasmid and phagemid cloning/ sequencing vectors. 

Plasmid Vectors for Facile Introduction of Passenger DNA and Selection of Recombinants. The map of a commonly used plasmid vector, pUC19 (7), is shown in Figure 2. Three parts of the vector are key to its utility. The origin sequence, oh, allows the repHcation of plasmid DNA in high copy number relative to the chromosome. A gene, amp, encoding the enzyme beta-lactamase, which hydrolyzes penicillin compounds, allows... 

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