Cloning and expression profiling of a cuticular protein gene in Daphnia carinata
Cloning and expression profiling of a cuticular protein gene in Daphnia carinata
复制标题
水蚤角质层蛋白基因的克隆和表达谱
DOI：
10.1007/s00427-014-0469-9
复制
发表时间：
2014-06-01
影响因子：
2.4
通讯作者：
Zhao, Yunlong
中科院分区：
文献类型：
--
作者：
Liu, Ajing;Zhang, Mingqing;Zhao, Yunlong
隆线水蚤对环境刺激的反应是从无性生殖到有性生殖的一个不寻常的转变。此前，D。在EST库中鉴定了隆线表皮蛋白（CP）。本研究克隆并测序了该基因的全长cDNA（GenBank登录号：KF 551931），并分析了其在不同繁殖状态下的表达水平。孤雌生殖和有性生殖的雌性D.采用半定量逆转录聚合酶链反应（RT-PCR）技术研究CP的表达。CP在两个生殖阶段都有表达，但在有性生殖的雌性中表达更高。还使用地高辛标记的RNA探针在RNA整体原位杂交试验中研究了细胞定位，CP主要表达在单性生殖生物的第一对胸附肢、头部表面、壳棘和表皮的其他部分。相反，CP的表达仅限于性雌性动物的胸附件。
The cladoceran Daphnia carinata undergoes an unusual transition from asexual to sexual reproduction in response to environmental stimuli. Previously, a D. carinata cuticular protein (CP) was identified in an EST library. In this study, the full-length CP cDNA was cloned and sequenced (GenBank accession number: KF551931), and the expression levels in different reproductive states were assessed. Parthenogenetic and sexual female D. carinata were isolated, and CP expression was investigated using semiquantitative reverse transcription polymerase chain reaction (RT-PCR). CP was expressed during both reproductive stages, but expression was higher in sexual females. Cellular localization was also investigated using digoxin-labeled RNA probes in RNA whole-mount in situ hybridization assays, and CP was mainly expressed in the first pair of thoracic appendages, the surface of the head, shell spines, and other parts of the epidermis in parthenogenetic organisms. In contrast, CP expression was restricted to the thoracic appendages in sexual females.