Explore the vast range of titles available.

S899.1; Objective]To obtain the full length cDNA sequence of the wax synthase (ws)gene of the white wax in-sect,Ericerus pela Chavannes,and heterologously express the enzyme in Escherichia coli.[Method]The 3′and 5′ends of the ws gene were obtained separately by using RACE,and the prokaryotic expression vector was constructed after the analysis of the full length cDNA sequence of the ws gene.Finally,the expression of WS in Ericerus coli BL21 was induced by IPTG.[Result]Sequence analysis showed that,the full-length cDNA of the ws gene was 1 518 bp,which included the 5′-UTR (untranslated region)with 94 bp,3′-UTR with 68 bp,and the open reading frame with 1 356 bp.The gene was deduced to encode 452 amino acid residues with the putative protein molecular weight of 51.8 kDa,and the theoretical isoelectric point of 6.35.Western Blot result showed that,the WS enzyme was expressed successfully in Ericerus coli.[Conclusion]In this study,the full length cDNA of the ws gene of Ericerus pela was obtained,and this enzyme w

Tropomyosin （TM） plays a critical role in skeletal and cardiac muscle development and function. To assess the fimctional significance of a-TM in Japanese flounder （Paralichthys olivaceus） development and metamorphosis, cDNA from Japanese flounder was cloned and a-TM mRNA measured during development and metamorphosis. The full-length cDNA is 1 191 bp, including a 5＇- untranslated region of 114 bp, a Y-UTR of 222 bp, and an open reading frame of 855 bp encoding a polypeptide of 284 amino acids. Real-time quantitative PCR revealed that a-TM mRNA is initially expressed in unfertilized ovum, indicating the a-TM gene is maternal. Relatively low mRNA levels were observed in different embryonic stages. A higher level of a-TM mRNA was detected 3 days post hatching （dph）, while the highest level was measured at 29 dph （metamorphic climax） after which it declined towards the end of metamorphosis. The expression of a-TM mRNA was up-regulated in thyroid hormone-treated larvae at 36 dph, but there was no marked difference

Cathepsin D （CTSD） is a lysosomal acidic endoproteinase that plays an important role in immune response, In this study, we obtained sweetfish （Plecoglossus altivelis） CTSD （PaCTSD） via de-novo transcriptome sequencing of sweetfish macrophages. The full length cDNA sequence of PaCTSD was 1955 bp encoding a propeptide of 397 amino acids. The deduced protein had a calculated molecular weight of 43.17x 103. Multiple alignment with other known CTSD amino acid sequences revealed amino acid conservation through the teleosts. Phylogenetic tree analysis showed that PaCTSD grouped tightly with other fish CTSD, and was close to that of Atlantic salmon and rainbow trout. Subsequently, PaCTSD was prokaryotically expressed and refolded by the urea gradient method on a nickel-nitrilotriacetic acid column. Enzyme activity analysis showed that PaCTSD exhibited pH-dependent proteolytic activity. Quantitative real-time PCR showed that PaCTSD mRNA was expressed in all detected tissues in healthy sweetfish. The highest expression

The signaling molecule hedgehog（Hh） is essential for cellular signaling required for body pattern formation in embryonic and individual development in vertebrate and invertebrates.Patched 1（Ptc1）,a receptor of Hh,mediates the activity of Hh by binging the Hh ligands.In this study,the Ptc1 of half-smooth tongue sole Cynoglossus semilaevis（CsPtc1） was cloned and characterized.The full-length of CsPtc1 cDNA is 5 212 nucleotides（nt）,and encodes a protein with 1 543 amino acid residues.CsPtc1 shared many features with Ptc1 of other telesot species.Real-time quantitative PCR revealed CsPtc1 mRNA expressed in brain,liver,heart,gill,intestines,spleen,gonad and kidney,with expression level in testis significantly higher than in ovary.In testis,hybridization signals were mainly detected in primary spermatocytes,secondary spermatocytes,and sertoli cells,while weak signals were found in oocytes.Correspondingly,the degree of methylation is higher in female than in male and pseudomale.Results indicate that CsPtc1 may be involved in Desert Hedgehog（DHH） maintenance of the male and pseudo-male germ line and spermatogenesis.

Harmful algal blooms （HABs） can occur and then disappear quickly, corresponding to consistent growing and declining of heavy biomasses. The molecular mechanism of blooming remains unclear. In this study, calmodulin gene （cam） of HAB causing species Alexandrium catenella was isolated and characterized, The expression of calmodulin gene was profiled at different growth rates and in heat stress. The full cDNA of cam was 597 nucleotides （nt） in length, including a 25 nt 5＇ untranslated region （UTR）, an 122nt 3＇ UTR, and a 450nt open reading frame （ORF） encoding 149 amino acids. The deduced calmodulin （CAM） was highly conserved in comparison with those of other organisms. As was determined with real-time RT PCR, the abundance of cam transcript varied in a pattern similar to cell growth rate during the whole growing period. The abundance of cam transcript increased by more than 8 folds from lag growth phase to exponential growth phase, and then obviously decreased from exponential growth phase to stationary/decline growth phase. In addition, the relative abundance of cam transcript significantly declined with time during heat shock. Taking CaM function described in other organisms into account, we believe that Ca2- -involved signal transduction, methyla- tion of DNA and toxin precursors underlined the cell growth of this species. The response of cam gene to heat stress in dinoflagellate suggested restrictions in Ca2＋ signal transduction and methylation. These findings are helpful to understand the relationships among growth, cell signal transduction, bloom formation and interaction with environmental stimuli in dinoflagellates.