Всероссийский научно-исследовательский институт физиологии, биохимии и питания животных – филиал Федерального государственного бюджетного научного учреждения «Федеральный научный центр животноводства – ВИЖ имени академика Л.К. Эрнста»
The object was to produce bovine and rabbit fibroblasts transformed by gene engineering construction with reporter gene of RFP, aimed to their use to produce transgenic animals by cloning technology. There are generated the culturies of fetal bovine and rabbit fibroblasts transformed by construction with gene of human lactoferrin (hLF) under promoter of bovine αS1-casein, reporter gene of RFP under cytomegalovirus (cmv) and selective gene of neomycin phosphotransferase (Neo) under promoter of virus sv40 (αS1-Cn-hLf-cmvRFP-svNeo). There are indicated that fetal rabbit fibroblasts are worse sensitive to transfection by Ca-phosphate method in comparison to bovine fibroblasts under adsorption time 24 hours and expression time 48 hours. Effective transfection of fetal rabbit fibroblasts was performed only under their treating by DMSO. Bovine fibroblasts have been effectively transformed without using protectors; transformation frequency was 8.6 per 105 cells and transformation efficiency was 3.5 per 105 cells and per 1 mg DNA of construction, under the use of 204 mg DNA and 7.6 mg DNA of carrier. Addition DNA-construction to DNA-carrier under fetal fibroblast transfection by lipofectamine method significantly increased transfection frequency of bovine fibroblasts and did not significantly effect the frequency transfection of rabbit fibroblasts. Transformed cells may be useful in nuclear transfer as a source of karyoplasts by reconstructed oocytes with the purpose for production of transgenic embryos and animals.
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