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Molecular cloning and mRNA expression of M-phase phosphoprotein 6 gene in black tiger shrimp (Penaeus monodon)
Jun Zhou1, Lihua Qiu, Shigui Jiang
1Biotechnology and Aquiculture Laboratory, The South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 231 Xingangxi Road, Guangzhou, 510300, People's Republic of China.
Abstract:
It is widely accepted that protein phosphorylation is a major control event in regulating cell cycle. In the present study, a novel M-phase phosphoprotein 6 (MPP6) was identified from black tiger shrimp Penaeus monodon (designated as PmMPP6) by cDNA library and RACE approaches. The full-length cDNA of PmMPP6 was of 690 bp, including a 5'-terminal un-translated region (5'UTR) of 68 bp, a 3'UTR of 172 bp with a poly (A) tail, and an open reading frame (ORF) of 450 bp encoding a polypeptide of 149 amino acids with a predicted molecular weight of 17.01 kDa. Blastx and phylogenetic analysis together supported that PmMPP6 was a novel member of shrimp MPP6. The mRNA expression of PmMPP6 in thirteen tissues was examined by real-time PCR, and mRNA transcript of PmMPP6 was predominantly detectable in tissues of lymphoid and muscle, to a lesser degree in the tissues of gill, ovary and hepatopancreas, and mainly detected in haemocytes, heart and gonad. The temporal expression of PmMPP6 in different developmental stages of ovary was investigated by real-time PCR. During the six stages of ovary development, two peaks expression of PmMPP6 was detected in stage II with 3.78-fold increase and stage V with 3.48-fold increase compared to that in stage I. All these results indicated that PmMPP6 might be involved in regulating shrimp cell cycle and ovary development.
Insights
Researchers identified a new M-phase phosphoprotein 6 (MPP6) in shrimp. This protein, PmMPP6, is predominantly found in lymphoid and muscle tissues and may regulate shrimp cell cycle and ovary development.
Area of Science:
- Marine Biology
- Molecular Biology
- Cell Biology
Background:
- Protein phosphorylation is crucial for cell cycle regulation.
- M-phase phosphoproteins (MPPs) play key roles in cell division.
Purpose of the Study:
- To identify and characterize a novel M-phase phosphoprotein 6 (MPP6) in the black tiger shrimp, Penaeus monodon.
- To investigate the expression patterns of this novel protein in various tissues and developmental stages.
Main Methods:
- cDNA library construction and RACE (Rapid Amplification of cDNA Ends) techniques were used for gene identification.
- Real-time PCR was employed to analyze mRNA expression levels in different tissues and during ovary development.
- Bioinformatic analyses including Blastx and phylogenetic analysis were performed.
Main Results:
- A novel gene, designated PmMPP6, was identified with a full-length cDNA of 690 bp encoding a 149-amino acid polypeptide.
- PmMPP6 mRNA was predominantly expressed in lymphoid and muscle tissues, with lower levels in gill, ovary, hepatopancreas, haemocytes, heart, and gonad.
- Two significant expression peaks of PmMPP6 were observed during ovary development (stage II and stage V).
Conclusions:
- PmMPP6 represents a novel shrimp MPP6 and is likely involved in regulating the cell cycle in Penaeus monodon.
- The distinct expression patterns suggest a role for PmMPP6 in shrimp ovary development.
