Authors : Raquel Cossio-Bayugar and Estefan Miranda-Miranda
Abstract: Rhipicephalus microplus in vitro cell culture have been useful in the study of acaricide resistance in ticks, a methodology for heterologous gene transference into these cultures may help to study several aspect of the cell biology and gene expression associated with acaricide resistance. In this study we assessed a pantropic retroviral gene transfer system in R. microplus embryonic cell cultures, for this purpose a Green Fluorescent Protein (GFP) was cloned downstream of the Drosophila heat shock protein 70 promoter within the pLNHX vector, pLNHX-GFP was cotransfected with pVSV-G vector coding for retroviral envelope protein into the GP-293 retroviral packaging cell line. An embryonic R. microplus cell line was transfected with the pantropic virus and evaluated for expression of the protein by fluorescence microscopy to detect the GFP. Infected embryonic tick cells expressed a green fluorescent signal that was not present in uninfected cultured cells. Relative Fluorescent Units (R.F.U) were determined as GFP expression values from infected and uninfected cells with mean values ranking at 1774 and 143 R.F.U. respectively, which demonstrates a successful heterologous gene expression in R. microplus cell culture. Our results showed that recombinant retroviral system can be used to express exogenous genes in R microplus cell cultures.
Raquel Cossio-Bayugar and Estefan Miranda-Miranda , 2007. Heterologus Gene Expression in a Cattle Tick Rhipicephalus microplus Embryonic Cell Culture . Journal of Animal and Veterinary Advances, 6: 1214-1218.