Phylogenetic tree of <i>lepr</i> amino acid sequences in four vertebrates and 22 invertebrates using the neighbour-joining method.

<p>The numbers in the phylogram nodes indicate percent bootstrap support for the phylogeny. The bar at the bottom indicates 5% amino acid divergence in sequences.</p

Nucleotide sequence and deduced amino acid sequence of Chinese mitten crab <i>lepr</i>.

<p>Nucleotides (upper) are numbered beginning at the 5′ end. Amino acids (lower), shown using one-letter abbreviations, are numbered beginning at the initiating methionine. The signal peptide is underlined, and the mature peptide is enclosed in the black box. The three conserved cysteine residues in the deduced amino acid sequence are shown in grey boxes. The stop codon is marked by an asterisk. The polyadenylation signal (AATAAA) is enclosed in the black ellipse. Arrows indicate the positions of primers; black boxes indicate the coding region; white lines indicate 5′ and 3′ UTRs; curly bracket indicate position of the EST clone. The <i>E. sinensis lepr</i> sequence was submitted to GenBank under accession number GU443952.</p

ClustalX alignment of <i>lepr</i> from <i>E. sinensis</i> and 25 other organisms.

<p>The vacuolar protein sorting 55 (Vps55) domain in <i>lepr</i> starts at Leu7 and ends at Asp127.</p

<i>Lepr</i> mRNA expression levels in six tissues from normal crabs (in the rapid developmental stage), precocious crabs and normal mature crabs.

<p>Values are shown as mean ± S.E. (n = 3). Significant differences relative to intestine are indicated with one asterisk (P<0.05) or with two asterisks (P<0.01).</p

<i>Lepr</i> sequences identities between <i>E. sinensis</i> and 25 other organisms.

<p><i>Lepr</i> sequences identities between <i>E. sinensis</i> and 25 other organisms.</p

Oligonucleotide primers used in the experiments.

<p>Oligonucleotide primers used in the experiments.</p

Tissue distribution of the <i>lepr</i> transcript measured by SYBR Green RT-PCR.

<p>Quantitative analysis of <i>lepr</i> gene expression relative to β-actin expression in different mitten crab tissues. Lepr gene expression was analyzed in the following tissues: In – intestine, TG – thoracic ganglia, Gn – gonad, AG – accessory gonad, Hp – hepatopancreas, CG – cranial ganglia, Ms – muscle, Gi – gill, Hr – heart, Hm – haemocytes and St – stomach. Expression data for each tissue were analyzed from three individual crabs. Vertical bars represent the mean ± S.E. (n = 3). Significant differences relative to controls are indicated with an asterisk (P<0.05) or with two asterisks (P<0.01).</p