Biopolym. Cell. 2007; 23(2):100-107.
Molecular and Cell Biotechnologies
Plasmid vector for the human preproinsuiin gene delivery into mammalian cells
1Toporova O. K., 1Kyrylenko S. D., 1Irodov D. M., 1Kordium V. A.
  1. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680

Abstract

Eukaryotic expression vector for human preproinsuiin gene has been constructed for further delivery into non-endocrine mammalian cells in vitro and in vivo for the development of experimental gene therapy for type I diabetes. Vector construction consists of two independent modules, namely, bacterial plasmid sequence that allows its replication in Escherichia coli cells, and expression cassette for the target and marker genes which is flanked by inverted terminal repeats of human adeno-associated virus. The human cytomegalovirus immediate-early promoter has been used to ensure the constitutive expression of human preproinsuiin gene. The hepatitis B viral enhancer 1 was subcloned into cassette to increase the target transgene expression. After vector construction in vitro transfer into mammalian cells of different origin, the stable transformants were obtained and the availability of target human preproinsuiin transgene in the transformed cells has been confirmed by PCR analysis.
Keywords: human preproinsuiin gene, expression vector, stable transformants

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