Molecular Cloning and Expression Analysis of Cyclin H from Black Tiger Shrimp (Penaeus monodon)

Abstract

Cyclin H is an important cell protein that plays a crucial role in cell division. In the present study, the cDNA of cyclin H (designated as Pmcyclin H) was identified from black tiger shrimp (Penaeus monodon) by expressed sequence tag (EST) analysis, and RACE techniques. The full length cDNA of Pmcyclin H is 1280bp, including a 5’-terminal un-translated region (5’UTR) of 63 bp, a 3’UTR of 218 bp with a poly (A) tail, and an open reading frame (ORF) of 999 bp encoding a polypeptide of 332 amino acids with a predicted molecular weight of 39 kDa, predicted pI of 6.39. Both Blast, and phylogenetic analysis, confirmed that Pmcyclin H is a new member of the shrimp cyclin H family. Using real-time PCR the mRNA expression of Pmcyclin H in eight tissues was examined, and mRNA transcript of Pmcyclin H was predominantly detectable in ovarian tissue, and to a lesser degree in the tissues of intestine, testis, stomach, and heart, but almost undetectable in the tissues of liver (hepar), brain, and muscle. The temporal expression of Pmcyclin H in different developmental stages of the ovaries was investigated by real-time PCR. During the six stages of ovarian development, one peak expression of Pmcyclin H was detected in stage II. All these results indicated that Pmcyclin H might be involved in the regulation of cell cycle and ovarian development of P. monodon.