Chymotrypsin and trypsin digested His-CI fragments were analyzed by Tris-Tricine SDS-16

<p><b>Copyright information:</b></p><p>Taken from "Repressor of temperate mycobacteriophage L1 harbors a stable C-terminal domain and binds to different asymmetric operator DNAs with variable affinity"</p><p>http://www.virologyj.com/content/4/1/64</p><p>Virology Journal 2007;4():64-64.</p><p>Published online 28 Jun 2007</p><p>PMCID:PMC1934351.</p><p></p>5% PAGE followed by silver staining. Molecular masses (in kDa) of marker proteins are shown at the left side of gel. 'Chy' and 'Try' indicate chymotrypsin and trypsin, respectively whereas, a – h indicate intact repressor, different digested fragments of repressor, respectively. N-terminal ends of fragments c and h were sequenced. Western blotting analysis of chymotrypsin/trypsin digested His-CI fragments from 2 and 30 mins incubations by a standard procedure as indicated in Materials and method. Summary of proteolysis. The putative domains of CI and its amino acid residues involved in formation of hinge, NTD and CTD are indicated

Repressor of temperate mycobacteriophage L1 harbors a stable C-terminal domain and binds to different asymmetric operator DNAs with variable affinity-3

<p><b>Copyright information:</b></p><p>Taken from "Repressor of temperate mycobacteriophage L1 harbors a stable C-terminal domain and binds to different asymmetric operator DNAs with variable affinity"</p><p>http://www.virologyj.com/content/4/1/64</p><p>Virology Journal 2007;4():64-64.</p><p>Published online 28 Jun 2007</p><p>PMCID:PMC1934351.</p><p></p>estimated from inset gel shift assay pictures) versus CI concentration (0.05 – 0.45 μM and 0.1 – 0.8 μM CI with and , respectively) are shown. Nearly 0.1 nM labeled operator was used in each reaction. Plot of %operator bound versus time shows the kinetics of CI dissociation from and operators in presences of excess cold operator. The amount of operator bound in the shifted complex of the zero time aliquot was considered as 100%. Plot of log Keq versus 1/T shows equilibrium binding of CI to operator at temperatures ranging from 25° – 42°C. All curves/lines are best-fit curves/lines

Repressor of temperate mycobacteriophage L1 harbors a stable C-terminal domain and binds to different asymmetric operator DNAs with variable affinity-2

<p><b>Copyright information:</b></p><p>Taken from "Repressor of temperate mycobacteriophage L1 harbors a stable C-terminal domain and binds to different asymmetric operator DNAs with variable affinity"</p><p>http://www.virologyj.com/content/4/1/64</p><p>Virology Journal 2007;4():64-64.</p><p>Published online 28 Jun 2007</p><p>PMCID:PMC1934351.</p><p></p>as calibrated with BSA (66 kDa, I), ovalbumin (46 kDa, II), carbonic anhydrase (29 kDa, III), and lysozyme (14.4 kDa, IV). Molecular weights were plotted against /, where and denote elution volume and void volume respectively. Void volume of column was determined from elution of blue dextran. Glutaraldehyde (GCHO) cross-linking. Nearly 0.5 μM His-CI or CTD was cross-linked with 0.1% GCHO and samples were analyzed by SDS-10% PAGE. Protein bands were visualized by silver staining. Horizontal arrows denote dimeric His-CI and CTD species