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Recombinant vaccinia constructs encoding HCMV antigens IE-1 (Vacc.IE-1), gB (Vacc.gB) and a negative control vaccinia virus construct made by insertion of the pSC11 vector alone, which is negative for thymidine kinase (Vacc.TK−), have been previously described [13].

The plaA- strain was generated by transforming Ax2 cells with a gene disruption construct made by inserting the bsr cassette into the EcoRV site of the genomic region of the plaA ORF.

Alternatively HMGB1 was expressed in E. coli as a fusion protein with a N-terminal (His 6 tag, using a plasmid construct made by inserting the rat HMGB1 gene into expression vector pET15b [22].

To test in vitro whether the methylation of the P2 promoter affects IGF1 gene expression, we used a reporter gene construct made by inserting a fragment of IGF1 promotertextendinging from −1,014 to +64 bp of the TSS - thus containing 7 of the studied CGs - into a promoter-less CpG-free luciferase expression plasmid.

Our particular emphasis in pretargeting is the use of bispecific trimeric (three Fab′s) recombinant constructs made by a modular method of antibody and protein engineering of fusion molecules called dock and lock (DNL).

All constructs made by PCR were sequenced to verify their integrity.

All constructs made by PCR were sequence verified.

An alternative to those vessels could be tissue-engineered vascular constructs made by self-organized tissue sheets.

Some of the frame configurations included strut constructs, made by one or two malleable stainless steel bars connecting the linear and circular elements by hooks [ 13].

This is the first example of transcription through a purely synthetic analogue of a DNA backbone, and it suggests that DNA constructs made by solid-phase synthesis and click ligation can be used directly for protein expression.

Unfortunately, the size of the matrices that can be constructed makes checking for outliers by hand impractical.