藜科的极端盐生植物盐穗木(Halostachys caspica)的高盐胁迫抑制差减文库中有39%的功能未知蛋白(proteins with obscure features,POFs),利用亚细胞定位分析可以初步判断其可能的功能.将盐穗木的1个POF c DNA序列Hc UKPP的编码区构建至p...藜科的极端盐生植物盐穗木(Halostachys caspica)的高盐胁迫抑制差减文库中有39%的功能未知蛋白(proteins with obscure features,POFs),利用亚细胞定位分析可以初步判断其可能的功能.将盐穗木的1个POF c DNA序列Hc UKPP的编码区构建至p CAMBIA1301-GFP植物表达载体上,冻融法将重组质粒p CAMBIA1301-Hc UKPP-GFP转化农杆菌EH105A,利用花序浸染法将基因导入拟南芥,经潮霉素筛选获得T1代阳性幼苗.通过激光扫描共聚焦显微镜观察转基因拟南芥植株的根部细胞.结果显示,表达GFP蛋白的对照转基因植株中,绿色荧光在细胞核、细胞膜以及细胞质中均能检测到,而表达Hc UKPP-GFP融合蛋白的转基因植株中,绿色荧光只在细胞质膜上表达,说明Hc UKPP蛋白为细胞质膜相关蛋白.本研究为深入探讨盐穗木未知蛋白的功能奠定了基础.展开更多
A lack of the complete pig proteome has left a gap in our knowledge of the pig genome and has restricted the feasibility of using pigs as a biomedical model.In this study,we developed a tissue-based proteome map using...A lack of the complete pig proteome has left a gap in our knowledge of the pig genome and has restricted the feasibility of using pigs as a biomedical model.In this study,we developed a tissue-based proteome map using 34 major normal pig tissues.A total of 5841 unknown protein isoforms were identified and systematically characterized,including 2225 novel protein isoforms,669 protein isoforms from 460 genes symbolized beginning with LOC,and 2947 protein isoforms without clear NCBI annotation in the current pig reference genome.These newly identified protein isoforms were functionally annotated through profiling the pig transcriptome with high-throughput RNA sequencing of the same pig tissues,further improving the genome annotation of the corresponding protein-coding genes.Combining the well-annotated genes that have parallel expression pattern and subcellular witness,we predicted the tissue-related subcellular locations and potential functions for these unknown proteins.Finally,we mined 3081 orthologous genes for 52.7% of unknown protein isoforms across multiple species,referring to 68 KEGG pathways as well as 23 disease signaling pathways.These findings provide valuable insights and a rich resource for enhancing studies of pig genomics and biology,as well as biomedical model application to human medicine.展开更多
According to the sequence of a 352bp fragment-M17 from the suppression subtractive hybridization library of the mutant line Cr3529 of Brassica napus,primers were designed and a 1000bp upstream fragment was cloned by 5...According to the sequence of a 352bp fragment-M17 from the suppression subtractive hybridization library of the mutant line Cr3529 of Brassica napus,primers were designed and a 1000bp upstream fragment was cloned by 5’-RACE,which was joined with the original piece and composed a 1343bp sequence.According to the 3’ and 5’ ends sequences,a pair of primers were designed and the encoding region sequence of this unknown protein was amplified and cloned.Sequence analyzing showed that it had 80.1% identity with an unknown-protein-encoding gene from Arabidopsis thaliana.Swiss-model was used to predict the structure of the induced protein,and a potential transmembrane helix from site 46 Leu to 66 Phe was identified.GLOBE prediction of globularity showed that this protein possessed a compact globular domain.展开更多
未知功能结构域蛋白家族(domains of unknown function protein families,DUFs)是一大群并未表征功能的蛋白家族,其蛋白结构中包含至少一个高度保守的DUF结构域。在众多的DUFs蛋白家族中存在一些植物特有的DUFs蛋白家族,其参与调控植物...未知功能结构域蛋白家族(domains of unknown function protein families,DUFs)是一大群并未表征功能的蛋白家族,其蛋白结构中包含至少一个高度保守的DUF结构域。在众多的DUFs蛋白家族中存在一些植物特有的DUFs蛋白家族,其参与调控植物生长发育、植物对病虫害的防御反应和植物对非生物胁迫的应答反应等生物学过程。本文对近几年关于植物DUFs基因参与上述生物学功能方面的研究进行了综述,旨在为未知功能基因的挖掘及其调控机制的阐明提供基础资料。展开更多
基金financially supported by the National Natural Science Foundations of China (Grant No. 31661143013)the Jinxinnong Animal Science Development Foundation
文摘A lack of the complete pig proteome has left a gap in our knowledge of the pig genome and has restricted the feasibility of using pigs as a biomedical model.In this study,we developed a tissue-based proteome map using 34 major normal pig tissues.A total of 5841 unknown protein isoforms were identified and systematically characterized,including 2225 novel protein isoforms,669 protein isoforms from 460 genes symbolized beginning with LOC,and 2947 protein isoforms without clear NCBI annotation in the current pig reference genome.These newly identified protein isoforms were functionally annotated through profiling the pig transcriptome with high-throughput RNA sequencing of the same pig tissues,further improving the genome annotation of the corresponding protein-coding genes.Combining the well-annotated genes that have parallel expression pattern and subcellular witness,we predicted the tissue-related subcellular locations and potential functions for these unknown proteins.Finally,we mined 3081 orthologous genes for 52.7% of unknown protein isoforms across multiple species,referring to 68 KEGG pathways as well as 23 disease signaling pathways.These findings provide valuable insights and a rich resource for enhancing studies of pig genomics and biology,as well as biomedical model application to human medicine.
文摘According to the sequence of a 352bp fragment-M17 from the suppression subtractive hybridization library of the mutant line Cr3529 of Brassica napus,primers were designed and a 1000bp upstream fragment was cloned by 5’-RACE,which was joined with the original piece and composed a 1343bp sequence.According to the 3’ and 5’ ends sequences,a pair of primers were designed and the encoding region sequence of this unknown protein was amplified and cloned.Sequence analyzing showed that it had 80.1% identity with an unknown-protein-encoding gene from Arabidopsis thaliana.Swiss-model was used to predict the structure of the induced protein,and a potential transmembrane helix from site 46 Leu to 66 Phe was identified.GLOBE prediction of globularity showed that this protein possessed a compact globular domain.
文摘未知功能结构域蛋白家族(domains of unknown function protein families,DUFs)是一大群并未表征功能的蛋白家族,其蛋白结构中包含至少一个高度保守的DUF结构域。在众多的DUFs蛋白家族中存在一些植物特有的DUFs蛋白家族,其参与调控植物生长发育、植物对病虫害的防御反应和植物对非生物胁迫的应答反应等生物学过程。本文对近几年关于植物DUFs基因参与上述生物学功能方面的研究进行了综述,旨在为未知功能基因的挖掘及其调控机制的阐明提供基础资料。
