Μελέτη της μετάθεσης ενδογενών ρετροϊκών στοιχείων VL30 σε μετασχηματισμένα κύτταρα ποντικού

Abstract

ΤΑ ΣΤΟΙΧΕΙΑ VL30 ΑΝΗΚΟΥΝ ΣΕ ΜΙΑ ΟΙΚΟΓΕΝΕΙΑ ΡΕΤΡΟΙΚΩΝ ΣΤΟΙΧΕΙΩΝ ΤΗΣ ΟΙΚΟΓΕΝΕΙΑΣΤΩΝ ΡΕΤΡΟΤΡΑΝΣΠΟΖΟΝΙΩΝ. ΤΑ ΡΕΤΡΟΤΡΑΝΣΠΟΖΟΝΙΑ ΕΙΝΑΙ ΚΙΝΗΤΑ ΓΕΝΕΤΙΚΑ ΣΤΟΙΧΕΙΑ ΠΟΥ ΚΑΤΑ ΤΗΝ ΕΙΣΧΩΡΗΣΗ ΤΟΥΣ ΔΗΜΙΟΥΡΓΟΥΝ ΜΕΤΑΛΛΑΓΕΣ. Ο ΣΚΟΠΟΣ ΤΗΣ ΕΡΓΑΣΙΑΣ ΜΑΣ ΗΤΑΝ ΝΑ ΔΙΕΡΕΥΝΗΣΟΥΜΕ ΤΗΝ ΠΙΘΑΝΟΤΗΤΑ ΤΑ ΣΤΟΙΧΕΙΑ VL30 ΝΑ ΜΕΤΑΤΙΘΕΝΤΑΙ ΣΕ ΚΥΤΤΑΡΑ NIH3T3 ΟΤΑΝ ΑΥΤΑ ΜΕΤΑΣΧΗΜΑΤΙΖΟΝΤΑΙ ΑΠΟ ΤΟΝ ΙΟ SV40.

THE VL30 ELEMENTS ARE A FAMILY OF DISPERSED RETROVIRUS LIKE DNA SEQUENCES CLASSIFIED IN THE VIRAL SUPERFAMILY OF RETROTRANSPOSONS. RETROTRANSPOSONS ARE MOBILE GENETIC ELEMENTS WHICH TRANSPOSE THROUGH AN RNA INTERMEDIATE AND CAN CREATE MUTATIONS WITH THEIR INSERTION. THE SCOPE OF OUR WORK WAS TO EXPLORE THE POSSIBILITY WHETHER VI30 ELEMENTS COULD TRANSPOSE IN CELLS TRANSFORMED BY SV40 VIRUS. WE CONSTRUCTED A RECOMBINANT VL30 VIRUS AND THEN ISOLATED CLONES WHICH CONTAINED ONE COPY OF THIS CONSTRUCT. THESE CLONES WERE INFECTED WITH SV40 VIRUS AND DERIVED TRANSFORMANTS WERE ISOLATED. THE DETECTION OF TRANSPOSITIONS WAS BASED ON COMPARISON OF RFLPS (RESTRICTION FRAGMENTS LENGTH POLYMORFISMS) BETWEEN THE PARENTAL AND SV40-TRANSFORMED CLONES. TRANSPOSITIONS WERE DETECTED AND THE NUMBERS WERE STATISTICALLY SIGNIFICANT. WE ALSO CARRIED OUT NESTED INVERSE PCR ANALYSIS ON DNA OF A TRANSPOSITION POSITIVE CLONE AND WE OBSERVED THE CHARACTERISTIC VIRAL INTEGRATION AND AN ACTUALTRANSPOSITION EVENT. WE INVESTIGATED THE EXPRESSION OF THE RECOMBINANT CONSTRUCT BY NORTHERN BLOT AND SI ANALYSIS AND THE RESULT SHOWED THAT THE TRANSPOSITION FREQUENTLY WASINDEPENDENT OF ITS LEVEL OF EXPRESSION.