针对浓香型白酒大曲样本,以16S r RNA基因为目的片段,分别采用16S r DNA克隆文库法和高通量测序法分析大曲中细菌微生物群落的组成,并通过丰度和多样性分析,比较了两种方法在研究大曲样品细菌多样性方面的适用性。结果表明,在门、纲、...针对浓香型白酒大曲样本,以16S r RNA基因为目的片段,分别采用16S r DNA克隆文库法和高通量测序法分析大曲中细菌微生物群落的组成,并通过丰度和多样性分析,比较了两种方法在研究大曲样品细菌多样性方面的适用性。结果表明,在门、纲、目、科和属的分类水平上,克隆文库方法检测大曲样本微生物得到4个门,4个纲,5个目,4个科,6个属;高通量测序得到13个门,22个纲,33个目,61个科,133个属。在门的水平上,克隆文库与高通量测序检测出优势类群的总数量与总丰度分别为3个(99.32%)和4个(98.61%),共有优势类群及其丰度分别为Firmicutes(88.88%和79.32%)、Proteobacteria(7.8%和15.04%)、Actinobacteria(2.72%和1.77%)。重复样本分析,得出的结果相似。克隆文库法与高通量测序法在反映样本微生物群落规模上差异较大,而在反应大曲样本中主要微生物的物种组成及数量比例上结果相近,特别是样本中优势微生物类群的结果基本相同。两种方法各具优势。展开更多
PA28α can activate the latent 20S proteasome together with PA28β,playing an important role in the processing of MHC class I antigen.PMSE1 gene encoding this activator has been characterized and documented in mammals...PA28α can activate the latent 20S proteasome together with PA28β,playing an important role in the processing of MHC class I antigen.PMSE1 gene encoding this activator has been characterized and documented in mammals,whereas,few reported among piscine.In the present study,two pairs of primer were designed and synthesised according to the full-length cDNA sequence of proteasome activator PA28α subunit which we had found in common carp.Using PCR two specific gene fragments of PA28α subunit were amplified from total genomic DNA extracted from the spleen of common,PCR products cloned into pMD18-T vector.The recombinant plasmids were verified by sequencing.The PA28α subunit gene(PSME1) of common carp had been successfully cloned.The sequence results were analysized with DNAStar,DNAMAN and BLAST software.Result indicated that carp PSME1 gene encompassed 3 602 nucleotides,11 exons,10 introns,which was very similar to the known PSME1 genes of other species with the same exon/intron arrangement.Three forms are shown at intron/exon boundaries of carp PSME1 gene,exon 5/intron 5 boundary belongs to class 1(GAA/G),exon 8/intron 8 boundary belongs to class 2(TCC/AA),the rest belong to class 0.The splice sites have been well conserved through evolution,and observe the regulation of GT-AG completely.A phylogenetic analysis using PA28α and PA28β protein sequences from the GenBank verifies the presumed orthologous relationships of the carp gene to their mammalian counterparts,and reveales a closer relationship between carp PA28α and zebrafish PA28α than between carp PA28α and mammalian PA28α.Comparing with human,pig,mouse,zebrafish,the structure of carp PMSE1 gene has been also well conserved through evolution.The base number of all exons is almost stable,althongh few introns(introns 1,5,7 in carp;introns 1, 4,7,8 in zebrafish) more variable than other three mammals,especially,the base number of intron 8 in zebrafish PSME1 gene.Our studies have demonstrated the carp PMSE1 gene,moreover,we have done a further work on the distinctions of PMSE1 genes among different species.However,further extensive study on this kinds of subunit genes will be necessary for more information about their molecular properties and functions.展开更多
文摘针对浓香型白酒大曲样本,以16S r RNA基因为目的片段,分别采用16S r DNA克隆文库法和高通量测序法分析大曲中细菌微生物群落的组成,并通过丰度和多样性分析,比较了两种方法在研究大曲样品细菌多样性方面的适用性。结果表明,在门、纲、目、科和属的分类水平上,克隆文库方法检测大曲样本微生物得到4个门,4个纲,5个目,4个科,6个属;高通量测序得到13个门,22个纲,33个目,61个科,133个属。在门的水平上,克隆文库与高通量测序检测出优势类群的总数量与总丰度分别为3个(99.32%)和4个(98.61%),共有优势类群及其丰度分别为Firmicutes(88.88%和79.32%)、Proteobacteria(7.8%和15.04%)、Actinobacteria(2.72%和1.77%)。重复样本分析,得出的结果相似。克隆文库法与高通量测序法在反映样本微生物群落规模上差异较大,而在反应大曲样本中主要微生物的物种组成及数量比例上结果相近,特别是样本中优势微生物类群的结果基本相同。两种方法各具优势。
文摘PA28α can activate the latent 20S proteasome together with PA28β,playing an important role in the processing of MHC class I antigen.PMSE1 gene encoding this activator has been characterized and documented in mammals,whereas,few reported among piscine.In the present study,two pairs of primer were designed and synthesised according to the full-length cDNA sequence of proteasome activator PA28α subunit which we had found in common carp.Using PCR two specific gene fragments of PA28α subunit were amplified from total genomic DNA extracted from the spleen of common,PCR products cloned into pMD18-T vector.The recombinant plasmids were verified by sequencing.The PA28α subunit gene(PSME1) of common carp had been successfully cloned.The sequence results were analysized with DNAStar,DNAMAN and BLAST software.Result indicated that carp PSME1 gene encompassed 3 602 nucleotides,11 exons,10 introns,which was very similar to the known PSME1 genes of other species with the same exon/intron arrangement.Three forms are shown at intron/exon boundaries of carp PSME1 gene,exon 5/intron 5 boundary belongs to class 1(GAA/G),exon 8/intron 8 boundary belongs to class 2(TCC/AA),the rest belong to class 0.The splice sites have been well conserved through evolution,and observe the regulation of GT-AG completely.A phylogenetic analysis using PA28α and PA28β protein sequences from the GenBank verifies the presumed orthologous relationships of the carp gene to their mammalian counterparts,and reveales a closer relationship between carp PA28α and zebrafish PA28α than between carp PA28α and mammalian PA28α.Comparing with human,pig,mouse,zebrafish,the structure of carp PMSE1 gene has been also well conserved through evolution.The base number of all exons is almost stable,althongh few introns(introns 1,5,7 in carp;introns 1, 4,7,8 in zebrafish) more variable than other three mammals,especially,the base number of intron 8 in zebrafish PSME1 gene.Our studies have demonstrated the carp PMSE1 gene,moreover,we have done a further work on the distinctions of PMSE1 genes among different species.However,further extensive study on this kinds of subunit genes will be necessary for more information about their molecular properties and functions.
