Molecular Cloning and Expression of Cryptocyanin Gene Isolated from an Indian Variety of Scylla olivacea.

Background: Molting and reproduction play vital roles in the life cycle of brachyuran crabs, and these two processes are closely interconnected. A key player in the molting cycle is cryptocyanin, which is similar to hemocyanin in sequence, size, and structure. Hemocyanin is a copper-containing oxygen-binding protein, while cryptocyanin is a copper-free protein that lacks oxygen-binding capacity.

Objective: The goal of the study was to carry out the isolation, cloning, and expression of the partial cryptocyanin gene from the Indian variety of Scylla olivacea.

Methods: The partial cryptocyanin gene was isolated from the hemocytes of the S. olivacea male and female crabs by qPCR for comparative expression analysis of the cryptocyanin gene.

Results: We successfully amplified, cloned, and expressed a 519bp partial cDNA encoding cryptocyanin from the Indian variety of Scylla olivacea, within the pRSET-B vector.

Conclusion: Our study showed that crustacean cDNA can be effectively expressed in bacterial vectors, and clones were stable for up to 6 months at -80oC. Real-time data showed a significant difference in cryptocyanin levels between male and female crabs. This finding highlights the need for further research with a larger sample size for better understanding.