Comparative analysis of inherited patterns of the transgene in transgenic mud loach Misgurnus mizolepis lines carrying the CAT reporter gene

Abstract

Comparative analyses of the inheritance patterns of transgene transmission and expression for up to F5 progenies were carried out in different lines of transgenic mud loach Misgurnus mizolepis containing the chloramphenicol acetyltransferase (CAT) reporter gene. Based on Southern blot hybridization, the analyses revealed different integration sites of transgenes in host chromosomes as well as different transmission modes among transgenic lines, including stable inheritance of integrated transgenic constructs, co-transmission of integrated and extrachromosomally persistant copies, and transgene rearrangement through successive generations. CAT-enzyme-linked immunosorbent assay expression analysis found three different major patterns of inherited expression of the transgene: (i) stable persistence of consistent level of expression; (ii) sudden loss of transgene expression following passage through the germline; and (iii) different levels of reduction in the expression in successive generations.