A Novel Approach of Transducing Recombinant Baculovirus into Primary Lymphoid Cells of Penaeus monodon for Developing Continuous Cell Line

Cell line development from shrimp is not a novel venture as researchers across the globe have been trying to have crustacean cell lines over 30 years. The reason for not attaining a crustacean or precisely a shrimp cell line is believed to be the replicative senescence and the inability to maintain...

Full description

Saved in:
Bibliographic Details
Published in:Marine biotechnology (New York, N.Y.) N.Y.), 2021-08, Vol.23 (4), p.517-528
Main Authors: B.S., Anoop, Puthumana, Jayesh, Sukumaran, Vrinda, Vazhappilly, Cijo George, Kombiyil, Salini, Philip, Rosamma, Singh, Isaac Sarojini Bright
Format: Article
Language:English
Subjects:
Aquatic crustaceans
Baculovirus
Biomedical and Life Sciences
Cell culture
Cell lines
Crustaceans
Culture media
Engineering
Freshwater & Marine Ecology
H-Ras protein
Immortalization
Life Sciences
Lymphoid cells
Marine crustaceans
Microbiology
Original Article
Penaeus monodon
Promoters
Recombinants
Senescence
Sodium
Sodium butyrate
Telomeres
Transduction
Zoology
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Cell line development from shrimp is not a novel venture as researchers across the globe have been trying to have crustacean cell lines over 30 years. The reason for not attaining a crustacean or precisely a shrimp cell line is believed to be the replicative senescence and the inability to maintain telomere length in vitro. Moreover, spontaneous in vitro transformations do not happen in shrimp cells. Oncogenic induction in primary cell culture is one of the ways to attain in vitro transformation by way of disrupting the mechanisms which involve cellular senescence. In this context, a recombinant baculovirus with shrimp viral promoter IHHNV-P2 was used for the transduction aimed at immortalization. An oncogene, H- ras , was successfully amplified and cloned in to the baculoviral vector, downstream to shrimp viral promoter IHHNV-P2 and upstream to GFP. Recombinant baculovirus with H- ras was generated and used for transduction into shrimp lymphoid cells during early dividing stage. Accordingly, fibroblast-like primary cell culture got developed, and H- ras and GFP expression could be confirmed. The study suggests that the simple method of incubating recombinant baculovirus with minced tissue enables in vitro transduction during early dividing stage of the cells, and the transduction efficiency gets enhanced by adding 5 mM sodium butyrate to the culture medium.
ISSN:1436-2228
1436-2236
DOI:10.1007/s10126-021-10043-6