Reactivity of anti-CsTKD1 mouse immune serum to <i>C. sinensis</i> CsTK D1D2 and recombinant CsTK D1 proteins by immunoblotting.

<p>Reactivity of anti-CsTKD1 mouse immune serum to <i>C. sinensis</i> CsTK D1D2 and recombinant CsTK D1 proteins by immunoblotting.</p

Purification of recombinant CsTKD1 by on-bead cleavage.

<p>Cs28GST-CsTKD1 was loaded to a glutathione sepharose 4B column and cleaved with thrombin. The cleaved-off CsTKD1 was eluted with PBS.</p

Developmental mRNA level of CsTK in <i>C. sinensis</i>.

<p>mRNA level was measured by quantitative real time PCR using a gene-specific primer pair.</p

Comparison of kinetic parameters of <i>C.sinensis</i> mutant TKs^*.

*<p>Value: mean of three assays ± SD.</p><p>NA, data not available.</p

Neighbor-Joining tree for the amino acid sequences of phosphagen kinases using a program in MEGA version 5.

<p>Bootstrap values were shown at the branching point (1,000 replications). Amino acid sequences were retrieved from DDBJ and GenBank.</p

Sequence identity of <i>C. sinensis</i> TK D1 and D2 domains to other phosphagen kinases^*.

*<p>Identity was calculated using ClustalW2 (<a href="http://www.ebi.ac.uk/Tools/msa/clustalw2/" target="_blank">http://www.ebi.ac.uk/Tools/msa/clustalw2/</a>).</p

Comparison intron/exon organization of <i>C. sinensis</i> TK with other phosphagen kinases.

<p>Intron positions of <i>C. sinensis</i> TK were based on aligned amino acid sequences. Nomenclature for the intron positions was taken from Uda et al. (2006) and Jarilla (2010). Intron phase is indicated by “.0”, “.1”, or “.2” following the amino acid sequence position. Conserved introns are shown by vertical dashes.</p

Enzyme activity of <i>C. sinensis</i> phosphagen kinase for various guanidine derivatives.

<p>Enzyme activity of <i>C. sinensis</i> phosphagen kinase for various guanidine derivatives.</p

Localization of CsTK in <i>C. sinensis</i> adults by immunohistochemical staining.

<p>Upper panels A–D were stained with anti-CsTKD1 mouse sera, and lower panels E-H with normal mouse serum. Mouse anti-CsTKD1 and normal sera were used at 1∶100 dilution. Panels A and E are testis (Ts); B and F, ventral sucker (VS); C and G, intestine (Int) with content in full; D and H, Testis and dreg (Dg) between two flukes. Tg, tegument; Ab, acini of biliary epithelium. Scale bar = 50 µm.</p

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