A small protein from the intergenic region of contains a zinc finger motif and regulates and transcription-3

<p><b>Copyright information:</b></p><p>Taken from "A small protein from the intergenic region of contains a zinc finger motif and regulates and transcription"</p><p></p><p>Molecular Microbiology 2008;67(4):772-780.</p><p>Published online Jan 2008</p><p>PMCID:PMC2253796.</p><p>© 2008 The Authors Journal compilation © 2008 Blackwell Publishing Ltd</p

A small protein from the intergenic region of contains a zinc finger motif and regulates and transcription-0

<p><b>Copyright information:</b></p><p>Taken from "A small protein from the intergenic region of contains a zinc finger motif and regulates and transcription"</p><p></p><p>Molecular Microbiology 2008;67(4):772-780.</p><p>Published online Jan 2008</p><p>PMCID:PMC2253796.</p><p>© 2008 The Authors Journal compilation © 2008 Blackwell Publishing Ltd</p

A selection of proteins containing a CPxCG-related zinc finger motif and definition of patterns and motifs to detect them

<p><b>Copyright information:</b></p><p>Taken from "A small protein from the intergenic region of contains a zinc finger motif and regulates and transcription"</p><p></p><p>Molecular Microbiology 2008;67(4):772-780.</p><p>Published online Jan 2008</p><p>PMCID:PMC2253796.</p><p>© 2008 The Authors Journal compilation © 2008 Blackwell Publishing Ltd</p> A large set of proteins shorter than 100 amino acids contain a CPxCG-related zinc finger motif. Several examples from (HS), (AF), (PA) and (TP) are listed. The upper part shows hypothetical and conserved hypothetical proteins which are unrelated to each of them outside the zinc finger patterns. The proteins in the lower part are functionally characterized. The zinc finger patterns are indicated by bold underlining. A definition of patterns and motifs is provided below the sequences. A CPxCG-related zinc finger motif consists of a pair of patterns, separated by 7–40 amino acids. One of the patterns must be a CPxCG-like pattern (grey), of which three forms exist. The other can be a more general Cys/His pattern containing two Cys or His residues separated by two or three intermediate amino acids (white). All eight types of the general Cys/His pattern are shown, the CPxCG-like pattern being a specification of one of them (CxxCx)

Biodistribution experiments of ¹²⁵I-L19-UG in F9 tumor-bearing mice.

<p>The percentage of the injected dose per gram of tissue (%ID/g) ± SD in the different organs, at the indicated time post ¹²⁵I-L19-UG i.v. administration, are reported.</p

Uteroglobin platform.

<p>A) The cDNA encoding the variable heavy (VH) and variable light (VL) domains of immunoglobulins composing a single chain fragment variable (scFv) is fused to the 5′ end of uteroglobin (UG) cDNA. The covalent dimerization of the UG moiety within the fusion protein scFv-UG allows the generation of a divalent antibody. B) The cDNA encoding for an scFv is fused to both the 5′ and 3′ ends of UG. The resulting fusion protein is a tetravalent antibody. C) Two different cDNAs encoding for two different antibody fragments or an antibody fragment and a therapeutic molecule, such as a cytokine, are fused to the 5′ and 3′ ends of UG. The resulting protein is dual specific and tetravalent <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0082878#pone.0082878-Ventura1" target="_blank">[26]</a>.</p

Mass spectrometry analysis of L19-UG from mammalian cells and <i>E.coli.</i>

<p>Mass spectrometry analysis of reduced L19-UG obtained from CHO cells (<b>A</b>, <b>C</b>) and from <i>E. coli</i> (<b>B</b>, <b>D</b>). The raw (<b>A</b>–<b>B</b>) and deconvoluted (<b>C</b>–<b>D</b>) mass spectra relative to the chromatographic peaks at 32.5 minutes are reported. The calculated average neutral mass of monomeric form of L19-UG is 34670,3 Da for the protein produced in CHO cells and 34670,6 Da for the protein produced in <i>E. coli</i>.</p

Immunoreactivity of L19-UG from <i>E.coli</i>.

<p><b>A)</b> Different concentrations of L19-UG purified from <i>E. coli</i> (black circles) and CHO cells (black diamonds) were tested for binding to the recombinant fragment 7.ED-B.8.9 FN by ELISA. To verify the specificity of the binding, different concentrations of L19-UG from <i>E. coli</i> were pre-incubated with a molar excess (1,2 milligrams per ml) of the recombinant ED-B and then tested in ELISA for its binding to 7.ED-B.8.9 (white circles). The mean absorbances at λ = 405 nm ± SD are indicated. <b>B)</b> A shift in the column retention volume of L19-UG in SEC (Superdex 200) was obtained after incubating L19-UG with a molar excess of the recombinant FN fragment B-8. Left panel: SEC profile of L19-UG showing a single peak at 15.14 ml. Right panel: SEC profile of L19-UG pre-incubated with a molar excess of B-8 rFN showing an elution peak at 13.65 ml, which corresponds to the immunocomplex L19-UG/B-8 rFN, and an elution peak at 16 ml, which corresponds to the excess unbound B-8 rFN.</p

L19-UG from <i>E. coli</i>.

<p><b>A)</b> Schematic of the L19-UG cDNA cloned into the pHEN-1 prokaryotic expression vector and fused to the pelB leader sequence. Depicted on the right is a schematic representation of the resulting dimeric fusion protein. <b>B)</b> SDS-PAGE analysis of L19-UG under non-reducing conditions after the first (lane 2) and the second purification steps (lane 3). L19-UG after protein refolding under non-reducing (lane 4) and reducing conditions (lane 5). Lane 1 shows the molecular mass standards. <b>C)</b> Size-exclusion chromatography (Superdex200 column) profiles of L19-UG purified from CHO cells (left panel) and from <i>E. coli</i> (central panel). The column retention volumes are 15.18 ml for the protein purified from CHO cells and 15.14 ml for the protein purified from <i>E. coli.</i> In the right panel the SEC profile of L19-UG obtained from <i>E. coli</i> after protein lyophilization and reconstitution. On the right the SDS-PAGE analysis of L19-UG, under non-reducing conditions, before (lane 1) and after (lane 2) protein lyophilization and reconstitution.</p

Maximum eGFP yield for all endogenous Sf21 promoters and early viral promoters in Hi5 cells.

<p>Maximum eGFP yield for all endogenous Sf21 promoters and early viral promoters in Hi5 cells.</p

Comparison of transient plasmid based expression in Hi5 cells with the OpIE2 promoter and HEK293-6E cells with the CMV promoter.

<p>Comparison of transient plasmid based expression in Hi5 cells with the OpIE2 promoter and HEK293-6E cells with the CMV promoter.</p