Lo Ting Sze, Cui Zhaoxia, Mong Janice L Y, Wong Queenie W L, Chan Siu-Ming, Kwan Hoi Shan, Chu Ka Hou
Department of Biology, The Chinese University of Hong Kong, Shatin, Hong Kong, China.
Mar Biotechnol (NY). 2007 Jul-Aug;9(4):459-68. doi: 10.1007/s10126-007-9006-4. Epub 2007 May 10.
To understand the molecular events of ovarian development in penaeid shrimp, RNA arbitrarily primed polymerase chain reaction (RAP-PCR) was used to identify differentially expressed genes during ovarian maturation in Metapenaeus ensis. From a screening of 700 clones in a cDNA library of the shrimp ovary by the products of RAP-PCR of different maturation stages, 91 fragments with differentially expressed pattern as revealed by dot-blot hybridization were isolated and sequenced. Forty-two of these fragments show significant sequence similarity to known gene products and the differentially expressed pattern of 10 putative genes were further characterized via Northern hybridization. Putative glyceraldehyde-3-phosphate dehydrogenase and arginine kinase are related to provision of energy for active cellular function in oocyte development. Translationally controlled tumor protein, actin, and keratin are related to the organization of cytoskeleton to accomplish growth and development of oocytes. High mobility group protein DSP1, heat shock protein 70, and nucleoside diphosphate kinase may act as repressors before the onset of ovarian maturation. Peptidyl-prolyl cis-trans isomerase and glutathione peroxidase are related to the stabilization of proteins and oocytes. This study provides new insights on the molecular events in the ovarian development in the shrimp.
为了解对虾卵巢发育的分子事件,采用RNA任意引物聚合酶链反应(RAP-PCR)来鉴定凡纳滨对虾卵巢成熟过程中差异表达的基因。通过不同成熟阶段的RAP-PCR产物对虾卵巢cDNA文库中的700个克隆进行筛选,分离出91个经斑点杂交显示具有差异表达模式的片段并进行测序。其中42个片段与已知基因产物具有显著的序列相似性,并且通过Northern杂交进一步鉴定了10个推定基因的差异表达模式。推定的甘油醛-3-磷酸脱氢酶和精氨酸激酶与为卵母细胞发育中的活跃细胞功能提供能量有关。翻译控制肿瘤蛋白、肌动蛋白和角蛋白与细胞骨架的组织有关,以完成卵母细胞的生长和发育。高迁移率族蛋白DSP1、热休克蛋白70和核苷二磷酸激酶可能在卵巢成熟开始前起抑制作用。肽基脯氨酰顺反异构酶和谷胱甘肽过氧化物酶与蛋白质和卵母细胞的稳定有关。本研究为对虾卵巢发育的分子事件提供了新的见解。