Cloning, expression and cellular localization of Daphnia pulex senescence-associated protein, DpSAP

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• 作者：Ajing Liu ; Ling Kong ; Mingqing Zhang ; Donglei Wu ; Danli Wang ; Yunlong Zhao
• 关键词：SAP ; senescence-associated protein ; DpSAP ; Daphnia pulex SAP ; RACE ; rapid amplification of cDNA ends ; RT-PCR ; reverse transcription polymerase chain reaction ; PCR ; polymerase chain reaction ; ORF ; open reading frame ; pI ; isoelectric point ; MW ; molecular weight ; NJ ; neighbor-joining ; ROS ; reactive oxygen species ; SOD ; superoxide dismutase ; POD ; peroxidase ; CAT ; catalase ; MDA ; malondialdehyde ; SDGs ; senescence-downregulated genes ; SAGs ; senescence-associated genes
• 刊名：Gene
• 出版年：25 January, 2014
• 年：2014
• 卷：534
• 期：2
• 页码：424-430
• 全文大小：1575 K

文摘

Daphnia (water fleas) are small crustaceans that undergo an unusual switch from asexual to sexual reproduction that is dependent on environmental conditions. In this study, a senescence-associated protein (SAP) from the common freshwater species Daphnia pulex was cloned using primers based on homologous sequences and rapid amplification of cDNA ends (RACE). Real-time PCR was employed to quantify the expression of D. pulex SAP (DpSAP) in individual organisms. The role of DpSAP in the reproductive transformation was further investigated in both parthenogenetic and sexual females by using digoxin-labeled SAP RNA probes and RNA whole-mount in situ hybridization. DpSAP was more highly expressed in sexual females, indicating a role in growth and reproduction. Cellular localization studies using RNA whole-mount in situ hybridization showed specific expression in the second tentacle joints. These expression patterns suggest an important role for DpSAP in the reproductive transformation of D. pulex.