Gene-engineering construction with structural gene of human lactoferrin under control of regulatory elements of bovine -casein gene with reporter gene EGFP under cytomegalovirus promoter and its influence on viability of pre-implantation rabbit embryo
With the use of PCR-amplification the fragment of plasmid <i>pсEGFP</i> with gene of modified green fluorescent protein (<i>EGFP</i>) under cytomegalovirus (<i>cmv</i>) promoter and polyadenilation site (<i>PolyA site BGH) </i>was obtained. This fragment was cloned into intermediary vector <i>pGEMT.</i> The plasmid so obtained was named as <i>pGEMTcmvEGFP</i>. Isolated and purified fragment <i>cmv-EGFP-PolyA site BGH</i> was cloned into plasmid <i>pas1Lf </i>on site<i> Not</i>I. So, the plasmid was obtained, named as <i>pas1LfGr</i>, <i> </i>which contained gene-engineering construction with two various genes: lactoferrin gene under regulatory elements of bovine gene of <i>-</i>casein and gene of modified green fluorescent protein under cytomegalovirus promoter. The whole construction was excised with restrictase <i>Cla</i>I (<i>Bsu</i>15I) and purified. After micro injection into pronucleus of rabbit's zygote the fluorescence was revealed in 35.4% of embryos and only 3.2% of microinjected embryos developed to stage of blastocyst (1/31). The data obtained suggest that there was significant decrease in survivability of rabbit's zygotes after <i>in vitro</i> micro injection into pronucleus.