运用c DNA末端快速扩增技术(RACE)和巢式PCR技术克隆出铁皮石斛硫氧还蛋白超家族基因(Thioredoxin like 2,DoTrxL2)。序列分析表明DoTrxL2基因开放阅读框(ORF)长度为639 bp,编码213个氨基酸,编码产物含有一个硫氧还蛋白家族保守的CXXC...运用c DNA末端快速扩增技术(RACE)和巢式PCR技术克隆出铁皮石斛硫氧还蛋白超家族基因(Thioredoxin like 2,DoTrxL2)。序列分析表明DoTrxL2基因开放阅读框(ORF)长度为639 bp,编码213个氨基酸,编码产物含有一个硫氧还蛋白家族保守的CXXC功能结构域,通过两个半胱氨酸的双硫键得失电子,起到氧化还原作用。分子进化分析显示,DoTrxL2同小兰屿蝴蝶兰、石刁柏亲缘关系较近。qRT-PCR分析显示,DoTrxL2在铁皮石斛原球茎不同发育时期及不同组织中均有表达差异,其中在原球茎形成早期表达量较高,表明该基因对原球茎形成有着促进作用;而在种子和花中表达量高,表明该基因对植物组织的生殖发育具有重要调控作用。展开更多
Thioredoxin (Trx) proteins are involved in many biological processes especially the regulation of cellular redox homeostasis. In this study, two Trx cDNAs were cloned from the mussel Mytilus galloprovincialis using ra...Thioredoxin (Trx) proteins are involved in many biological processes especially the regulation of cellular redox homeostasis. In this study, two Trx cDNAs were cloned from the mussel Mytilus galloprovincialis using rapid amplification of cDNA ends-polymerase chain reaction (RACE-PCR). The two cDNAs were named MgTrx1 and MgTrx2, respectively. The open reading frames of MgTrx1 and MgTrx2 were 318 and 507 base pairs (bp) and they encoded proteins of 105 and 168 amino acids with estimated molecular masses of 11.45 and 18.93 kDa, respectively. Sequence analysis revealed that both proteins possessed the conserved active site dithiol motif Cys-Gly-Pro-Cys. In addition, MgTrx2 also possessed a putative mitochondrial targeting signal suggesting that it is located in the mitochondria. Quantitative real-time polymerase chain reaction (qPCR) revealed that both MgTrx1 and MgTrx2 were constitutively expressed in all tissues examined. The MgTrx1 transcript was most abundant in hemocytes and gills, whereas the MgTrx2 transcript was most abundant in gonad, hepatopancreas, gill and hemocytes. Following Vibrio anguillarum challenge, the expression of MgTrx1 was up-regulated and reached its peak, at a value 10-fold the initial value, at 24 h. Subsequently, expression returned back to the original level. In contrast, the expression level of MgTrx2 was down-regulated following bacterial stimulation, with one fifth of the control level evident at 12 h post challenge. These results suggest that MgTrx1 and MgTrx2 may play important roles in the response of M. galloprovincialis to bacterial challenge.展开更多
文摘运用c DNA末端快速扩增技术(RACE)和巢式PCR技术克隆出铁皮石斛硫氧还蛋白超家族基因(Thioredoxin like 2,DoTrxL2)。序列分析表明DoTrxL2基因开放阅读框(ORF)长度为639 bp,编码213个氨基酸,编码产物含有一个硫氧还蛋白家族保守的CXXC功能结构域,通过两个半胱氨酸的双硫键得失电子,起到氧化还原作用。分子进化分析显示,DoTrxL2同小兰屿蝴蝶兰、石刁柏亲缘关系较近。qRT-PCR分析显示,DoTrxL2在铁皮石斛原球茎不同发育时期及不同组织中均有表达差异,其中在原球茎形成早期表达量较高,表明该基因对原球茎形成有着促进作用;而在种子和花中表达量高,表明该基因对植物组织的生殖发育具有重要调控作用。
基金Supported by the National Natural Science Foundation of China(No.31172388)the 100 Talents Program of the Chinese Academy of Sciencesthe Key Laboratory for Ecological Environment in Coastal Areas(201011),State Oceanic Administration
文摘Thioredoxin (Trx) proteins are involved in many biological processes especially the regulation of cellular redox homeostasis. In this study, two Trx cDNAs were cloned from the mussel Mytilus galloprovincialis using rapid amplification of cDNA ends-polymerase chain reaction (RACE-PCR). The two cDNAs were named MgTrx1 and MgTrx2, respectively. The open reading frames of MgTrx1 and MgTrx2 were 318 and 507 base pairs (bp) and they encoded proteins of 105 and 168 amino acids with estimated molecular masses of 11.45 and 18.93 kDa, respectively. Sequence analysis revealed that both proteins possessed the conserved active site dithiol motif Cys-Gly-Pro-Cys. In addition, MgTrx2 also possessed a putative mitochondrial targeting signal suggesting that it is located in the mitochondria. Quantitative real-time polymerase chain reaction (qPCR) revealed that both MgTrx1 and MgTrx2 were constitutively expressed in all tissues examined. The MgTrx1 transcript was most abundant in hemocytes and gills, whereas the MgTrx2 transcript was most abundant in gonad, hepatopancreas, gill and hemocytes. Following Vibrio anguillarum challenge, the expression of MgTrx1 was up-regulated and reached its peak, at a value 10-fold the initial value, at 24 h. Subsequently, expression returned back to the original level. In contrast, the expression level of MgTrx2 was down-regulated following bacterial stimulation, with one fifth of the control level evident at 12 h post challenge. These results suggest that MgTrx1 and MgTrx2 may play important roles in the response of M. galloprovincialis to bacterial challenge.
