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The Chinese mitten crab Eriocheir sinensis is an important aquaculture species in Asia and causes considerable economic and ecological damage as an invasive species in North America and Europe. Until recently, molecular markers available for genetic analysis in this species were lim- ited to a few microsatellite sequences used in population studies. Here we describe the discovery of 3191 single nucleotide polymorphisms (SNPs) from the alignment of expressed sequence tags (ESTs) in E. sinensis. The observed frequency of SNPs was estimated at 0.78 per 100 bp of contig sequences. C/T substitutions were frequent and accounted for 32.2% of all SNPs. A subset of these SNPs (n = 38) was selected for validation by allele-specific PCR with melting temperature (Tm)-shift primers based on their observed frequency in sequence data; 12 (31.6%) of these were polymorphic in a panel of 40 wild-caught crabs. Eight (21.1%) did not amplify any product, and 18 (47.4%) failed due to amplifi- cation failure of 1 allele-specific primer. A table of optimal codons was deduced from the analysis of the E. sinensis EST dataset, and the results implied that the gene expression levels and GC-content might play important roles in codon usage bias. Sixteen codons ending with a G or C base were defined as 'optimal codons,' which may provide useful information for predicting gene expression in crabs. These are the first SNPs developed for the Chinese mitten crab and will provide a useful complement to currently available genetic markers.

The structure and organization of mitochondrial genomes of 14 marine bivalves and two freshwater bivalves were analyzed using comparative genomics and bioinformatics methods. The results showed that the organization and gene order of the mitochondrial genomes of these bivalve species studied were different from each other. The size, organization, gene numbers, and gene order of mitochondrial genomes in bivalves at different taxa were different. Phylogenetic analysis using the whole mitochondrial genomes and all the coding genes showed different results-- phylogenetic analysis conducted using the whole mitochondrial genomes was consistent with the existing classification and phylogenetic analysis conducted using all coding genes not consistent with the existing classification.

AFLP markers were used to construct the primary linkage map in a family of Mizuhopecten yessoensis. A total of 1 855 markers were generated in two parents and 52 progenies of the mapping family by using 56 AFLP primer combinations. Among the 1 855 markers, 598 were polymorphic and 354 were in agreement with the Mendelian segregating ratio of 1:1. Markers segregated according to Mendelian 1:1 ratio (P>0.05) and 23 distorted markers (0.01

MYP (Major yolk protein) gene expression at transcription level in different stages of gonad of sea urchin Strongylocentrotus intermedius and hybrids (S. intermediusfemale symbolxS. nudusmale symbol) was analyzed by real-time RT-PCR. Based on normalization with 18S rRNA levels, the comparative quantities of MYP expression were determined. The expression of MYP gene in gonad showed little difference between female and male. MYP gene expression was decreased rapidly in the gonad of S. intermedius at different stages, and slowly in hybrids. The comparative quantities of MYP expression in the gonads of S. intermedius were decreased from 44.55% to 9.59% in female and from 41.17% to 1.83% in male at different stages. The comparative quantities of MYP expression in the gonads of the hybrids were decreased from 37.66% to 19.22% in female and from 36.66% to 12.55% in male at different stages. The results indicated that the difference of MYP expression was correlated with the variation caused by hybridization.

The lipopolysaccharide-induced TNF-alpha factor (LITAF) is an inflammatory cytokine, which plays an important role in innate immunity system. Based on the expressed sequence tag (EST) of Japanese scallop (Mizuhopecten yessoensis), the cDNA of LITAF gene was amplified using rapid amplification of cDNA ends (RACE) approach. Results showed that the full-length cDNA of LITAF is 1 551 bp consisting of a 5' untranslated region (UTR) of 76 bp, a 3' UTR of 1 001 bp, and an open reading frame (ORF) of 474 bp encoding a polypeptide of 157 amino acids, and there is a conserved LITAF domain in amino acid sequences. The estimated molecular mass is 16.99 kDa and the theoretical isoelectric point is 6.24. The total length of LITAF is 3 698 bp, which includes three exons and two introns. Real-time quantitative PCR was carried out to measure LITAF mRNA expression in adult tissues and monitor mRNA expression patterns during embryonic development after bacteria (Vibrio anguillarum) challenged. The expression level of LITAF mRNA was detected in all the adult tissues with the highest in the kidneys. The trochophore owns the highest expression level of LITAF in embryonic development. LITAF expression showed significant difference(P0.01)between the control and bacteria challenged specimens at 36 h. These results suggest that the LITAF should be a member of the LITAF family that perhaps involved in the innate immune response of Japanese scallop.

The structure and organization of mitochondrial genomes of 14 marine bivalves and two freshwater bivalves were analyzed using comparative genomics and bioinformatics methods. The results showed that the organization and gene order of the mitochondrial genomes of these bivalve species studied were different from each other. The size, organization, gene numbers, and gene order of mitochondrial genomes in bivalves at different taxa were different. Phylogenetic analysis using the whole mitochondrial genomes and all the coding genes showed different results-- phylogenetic analysis conducted using the whole mitochondrial genomes was consistent with the existing classification and phylogenetic analysis conducted using all coding genes not consistent with the existing classification.

AFLP markers were used to construct the primary linkage map in a family of Mizuhopecten yessoensis. A total of 1 855 markers were generated in two parents and 52 progenies of the mapping family by using 56 AFLP primer combinations. Among the 1 855 markers, 598 were polymorphic and 354 were in agreement with the Mendelian segregating ratio of 1:1. Markers segregated according to Mendelian 1:1 ratio (P0.05) and 23 distorted markers (0.01P0.05) were used in linkage analysis. The male map was composed of 178 markers in 22 linkage groups, covering a total of 1 263.0 cM with the average interval of 8.1 cM between markers, and the observed coverage was 79.4%. The female map was composed of 163 markers in 22 linkage groups, covering a total of 1 259.5 cM with the average interval of 8.9 cM between markers, and the observed coverage was 76.8%. AFLP markers showed an uneven distribution on the linkage group and there were a lot of markers with a distance of 0.

MYP (Major yolk protein) gene expression at transcription level in different stages of gonad of sea urchin Strongylocentrotus intermedius and hybrids (S. intermediusfemale symbolxS. nudusmale symbol) was analyzed by real-time RT-PCR. Based on normalization with 18S rRNA levels, the comparative quantities of MYP expression were determined. The expression of MYP gene in gonad showed little difference between female and male. MYP gene expression was decreased rapidly in the gonad of S. intermedius at different stages, and slowly in hybrids. The comparative quantities of MYP expression in the gonads of S. intermedius were decreased from 44.55% to 9.59% in female and from 41.17% to 1.83% in male at different stages. The comparative quantities of MYP expression in the gonads of the hybrids were decreased from 37.66% to 19.22% in female and from 36.66% to 12.55% in male at different stages. The results indicated that the difference of MYP expression was correlated with the variation caused by hybridization.