Monoclonal Antibody Response, Sfil and Ecori Fragments of Genome DNA of Theileria Parva Melela of Eastern Tanzania

Abstract

Genomic DNA and monoclonal antibody (Mab) response of Theileria parva Melela, of Morogoro, Tanzania were compared with those of T.parva Muguga. Using indirect fluorescent antibody test. Positive reactions at 1:50 and 1:200 dilutions were observed with MAb 1, 2, 3,  4,  10, 12 and 22, nonspecific fluorescence in < 0.1% of cells for MAb 7 and 21 and negative for MAb 15, 20 and 23 with very close response to T. parva Muguga. There were restriction fragment length polymorphisms with Muguga stock on digestion of genomic DNA by EcoRI and Sfil endonucleases, separated by agarose and pulsed field gel electrophoresis respectively. T. parva Melela DNA digested with EcoRI resolved multiple restriction fragment bands 603- 2400 kb. Sfil digestion and pulsed field gel electrophoresis resolved 24 discrete fragments 65-1800 kb. There were more fragments above 800 kb than in T. parva Muguga, indicating a large genome of 1200-1400 megabase pairs (mb) for Melela T. parva compared to 10 mb for Muguga. Variations in hybridization signals for T. parva Melela and Muguga were observed when southern blots were probed with small subunit ribosomal RNA gene probe pRIBSSI, 1.4 kb repetitive T. parva sequence Tprl PMB3 probe cloned in plasmid pUC8, for EcoRI digested DNA and 8.1 kb synthetic telomeric oligonucleotide DNA probe for Sfil DNA digests.