Molecular cloning of cyclin B transcript with an unusually long 3?untranslation region and its expression analysis during oogenesis in the Chinese mitten crab, Eriocheir sinensis

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• 作者：Jun-Jiang Fang and Gao-Feng Qiu
• 关键词：Cyclin B ; cDNA cloning ; Differential expression ; Oogenesis ; 3′ ; untranslated regions
• 刊名：Molecular Biology Reports
• 出版年：2009
• 出版时间：July, 2009
• 年：2009
• 卷：36
• 期：6
• 页码：1521-1529
• 全文大小：509.1 KB

文摘

The meiotic maturation of oocyte in animals is regulated by maturation promotion factor (MPF), a complex of Cdc2 and cyclin B. Although the role of MPF during oocyte maturation has been well studied in a wide variety of eukaryotic organisms, little is known for crustacean species. In this study, a full-length cDNA of cyclin B was cloned from the Chinese mitten crab using degenerate RT-PCR and RACE methods. The crab cyclin B cDNA was 3,794 bp containing an unusually long 3′ untranslation region (UTR) of 2,403 bp and an open-reading frame encoding for a protein of 410 amino acids, with a calculated molecular mass of 45 kDa. The long 3′UTR harbors many cytoplasmic polyadenylation elements (CPE), and the GY-box, Brd-box, K-box that are perfectly complementary to the 5′-ends of various Drosophila microRNAs. The crab cyclin B transcript was predominantly expressed in ovary and testis. Semi-quantitative RT-PCR analysis revealed that the amount of cyclin B mRNA was high at previtellogenesis and late vitellogenesis stages, while low at early and middle vitellogenesis, suggesting that differential expression of cyclin B is closely related to oogonial proliferation (mitosis) and oocyte meiotic maturation.