Submitted to: In Vitro Cellular and Developmental Biology - Plants
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 11/10/2003
Publication Date: 9/30/2003
Citation: GRASELA, J.J., MCINTOSH, A.H. APPLICATION OF INTER-SIMPLE SEQUENCE REPEATS (ISSR) TO INSECT CELL LINES. IN VITRO CELLULAR AND DEVELOPMENTAL BIOLOGY - PLANTS. 39:353-363. 2003. Interpretive Summary: Insect cell lines are used in the production of baculoviruses, which are important biological control agents of many agricultural insect pests. Currently, there are no available techniques that can identify which tissue in the insect a particular cell line was derived from, nor is there a technique to distinguish between the parent cell line and a copy of itself. Using what are called inter-simple sequence repeat (ISSR) primers to generate a DNA pattern, we were able to easily distinguish between cell lines derived from different tissues of the same species, between copies of cells and their parental cells, and between cells from different species. By identifying and characterizing insect cells from different species, researchers are able to avoid cross-contamination of different cells and selectively determine the ideal cell line for optimal production of an insect virus. The cell line could then be further used in both laboratory studies and in the large-scale commercial production of the virus. This ISSR approach will provide researchers with an invaluable technique for identifying insect cell lines with potentially important ramifications for the laboratory production of baculovirus control agents.
Technical Abstract: Inter-simple sequence repeat (ISSR) primers designed to anneal to microsatellites were used to obtain DNA fingerprint profiles to distinguish between 16 established insect cell lines derived from an assortment of lepidopteran, dipteran, and coleopteran species. Three different levels of cell line comparison were made: (1) between parents and their clones; (2) between cell lines derived from different tissues from the same species; and (3) between cell lines derived from different insect species. Of the 16 repeat oligonucleotide primers employed in this study, nine primers generated several unique markers to distinguish between parental cell lines and their clones. Four of the 16 primers also generated DNA profiles with a number of unique bands, enabling the distinction between cell lines derived from specific tissues from the same species. In addition, ISSR-generated DNA profiles provided the greatest number of unique markers to easily distinguish between insect cell lines derived from different species.