Scientia Agricultura Sinica ›› 2014, Vol. 47 ›› Issue (13): 2659-2670.doi: 10.3864/j.issn.0578-1752.2014.13.018

• ANIMAL SCIENCE·VETERINARY SCIENCERE·SOURCE INSECT • Previous Articles     Next Articles

Identification, Phylogenetic and Tissue Expression Microarray Analyses of the Sirtuin Gene Family in Silkworm (Bombyx mori)

 CHEN  Cong, SONG  Jiang-Bo, MENG  Gang, TONG  Xiao-Ling, DAI  Fang-Yin, LU  Cheng   

  1. College of Biotechnology, Southwest University/State Key Laboratory of Silkworm Genome Biology/Key Laboratory for Sericulture Functional Genomics and Biotechnology of Ministry of Agriculture, Chongqing 400716
  • Received:2013-12-16 Online:2014-07-01 Published:2014-01-26

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Abstract: 【Objective】The objectives of this study are to identify sirtuin genes from silkworm genome, analyze the gene structure, protein structure, physicochemical properties, phylogeny and tissue expression pattern, and further to provide a basis for the function of sirtuins of silkworm and silkworm model organismal system.【Method】Based on silkworm genome database and bioinformatic method, silkworm sirtuin family genes were identified by comparative genomic analysis. The opening reading frame (ORF) of sirtuins was predicted by ORFfinder, exon and intron were predicted by SIM4. Gene structure was created using GSDS. Physicochemical properties of sirtuin proteins were done by ExPaSy server, multiple sequence alignment and its secondary structure analysis were done by CLUSTAL_X and ESpript, protein structure and phylogenetic tree were analyzed using SMART and MEGA5.0 software. The expression pattern of sirtuin genes in different tissues was analyzed based on the existing microarray database of 3-day-old 5th instar larvae. The expression level of sirtuin genes of silkworm in different tissues of 3-day-old 5th instar larvae were determined by quantitative RT-PCR.【Result】A total of 5 sirtuin genes (Bmsirt2, Bmsirt4, Bmsirt5, Bmsirt6, Bmsirt7) were systematically identified from silkworm and classified into 4 classes (I, II, III, IV). These genes were distributed on 5 chromosomes, and they were single copy genes. Homologous alignment and phylogenetic analysis revealed that the sirtuin genes in silkworm had apparent orthologous relationship with homologous genes from other insects respectively and they were highly homologous, also lack of sirt3. Protein structures showed that sirt6 had two sir2 domain associated together, as the same as other organisms. Microarray analysis showed that the sirtuin family genes had transcriptional active in multiple tissues. Analysis by real time quantitative PCR indicated that Bmsirt4 was expressed relatively lower abundance in fatboby and silk gland. Bmsirt5 was expressed relatively higher abundance in testis and midgut, but was lower abundance in fatboby, hemocyte, and silk gland. This was consistent with the microarray data. 【Conclusion】 Five genes of sirtuin family in silkworm were identified by genome-wide screening, they were classified into 4 classes, highly homologous with other insects. Microarray data was consistent with the result of real-time fluorescence quantitative PCR, and the analysis showed that the tissue expression pattern was diversity.

Key words: sirtuin , bioinformatics , expression characteristics , Bombyx mori

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