title: Molecular cloning and functional characterization of cyclin E and CDK2 from Penaeus monodon.
abstract: Reduced reproductive performance of the black tiger shrimp (Penaeus monodon) has caused economic losses and hampered the fishing industry. Detailed investigation of the molecular mechanism by which the cell cycle is regulated in this organism is needed to understand the development and maturation of ovaries and oocytes, with a view to improving reproductive capacity. Cell cycle progression is mainly determined by cyclin-dependent kinase (CDK) and cyclin complexes, the cyclin E/CDK2 complex playing a key role in G1/S transition. However, knowledge of the interplay between cyclin E and CDK2 in invertebrates remains limited. In this study, full-length P. monodon cyclin E (Pmcyclin E) and CDK2 (PmCDK2) sequences were cloned. The open reading frame of Pmcyclin E was 1263 bp in length and encoded a 47.9-kDa protein, while that of PmCDK2 was 921 bp, encoding a protein of 34.9 kDa. Recombinant cyclin E and CDK2 proteins were expressed in Escherichia coli and purified by Ni-chelating affinity chromatography. In addition, a pull-down assay was performed to identify any interaction between Pmcyclin E and PmCDK2. This research provides a basis for the study of the functional mechanisms of the cyclin E/CDK2 complex in shrimp, further enriching our knowledge of invertebrate cell cycle regulation.
authors.affiliation: South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, China.|Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture, Guangzhou, China.
authors.forename: C
authors.initials: C
authors.lastname: Zhao
authors.affiliation: South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, China.|Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture, Guangzhou, China.
authors.forename: M J
authors.initials: MJ
authors.lastname: Fu
authors.affiliation: South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, China qiugroup_bio@outlook.com.|Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture, Guangzhou, China qiugroup_bio@outlook.com.|Tropical Aquaculture Research and Development Center of South China Sea Fisheries Research Institute, Sanya, China qiugroup_bio@outlook.com.
authors.forename: L H
authors.initials: LH
authors.lastname: Qiu
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D019927:Cyclin E / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D051357:Cyclin-Dependent Kinase 2 / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D005786:Gene Expression Regulation; D033561:Penaeidae / Q000235:genetics*
chemical_list: D019927:Cyclin E; D051357:Cyclin-Dependent Kinase 2