Inheritance and expression stability of red fluorescent protein gene in transgenic (Tanichthys albonubes)

The stability of the red fluorescent protein (RFP) transgenes is important for application of the transgenic Tanichthys albonubes line.This paper investigated comparatively the inheritance and expression of RFP gene in different transgenic T. albonubes generations. Under the observation of fluorescence microscope, RFP gene was expressed in almost all body tissues, and both F6 and F10 generations had a similar expression pattern. Results of self-cross and test-cross breeding experiments of transgenic F6 and F10 individuals indicated that segregation of RFPgene still followed Mendelian single-gene inheritance, and their transgenic progenies had a uniform phenotype. The regions including myosin light polypeptide 2 promoter,RFPgene coding sequence and transgene flanking junctions were amplified by PCR from genomic DNA of F2, F6 and F10 generations. Sequence alignment analysis showed that these important regions were well preserved in transgenic fish, and any kind of alteration at nucleotide level was not found. These results suggested that, RFP gene in transgenic T. albonubes maintained stable inheritance and expression through successive generations.