Fluorescence in situ hybridization landmarks for chromosomes of Culicoides variipennis (Diptera: Ceratopogonidae)

Because the three chromosomes of Culicoides variipennis (Coquillett) are morphologically indistinguishable, physical landmarks were needed so that the chromosomes can be identified uniquely and oriented unambiguously before initiating the construction of a physical map based on FISH (fluorescence in situ hybridization). When repetitive sequence clones p1887 and K8.1a8 were labeled with digoxigenin and probe K8a.2G2 was labeled with biotin-11-dUTP and digoxigenin, the 3 chromosomes could be differentiated unambiguously when visualized with specific band-pass filters. This provided the basis for C. variipennis FISH landmark probes that enabled the identification and orientation of all 3 pairs of chromosomes. Using this multicolor FISH labeling strategy, probes that provide unique landmarks for the C, variipennis FISH physical map have been found and may be used in all FISH reactions where an unknown probe is placed to metaphase chromosomes of C. variipennis. A physical map of the C. variipennis genome will provide the foundation for map-based positional cloning of the gene(s) that control vector competence for the bluetongue viruses.