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

Abstract

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 F 6 and F 10 generations had a similar expression pattern.Results of self-cross and test-cross breeding experiments of transgenic F 6 and F 10 individuals indicated that segregation of RFP gene still followed Mendelian single-gene inheritance,and their transgenic progenies had a uniform phenotype.The regions including myosin light polypeptide 2 promoter,RFP gene coding sequence and transgene flanking junctions were amplified by PCR from genomic DNA of F 2,F 6 and F 10 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.