Gene therapy DNA vector VTvaf17, method of production; Escherichia coli strain SCS110-AF, method of production; Escherichia coli strain SCS110-AF/VTvaf17 bearing gene therapy DNA vector VTvaf17, method of production

摘要

Disclosed is a gene therapy DNA vector VTvaf17 for genetic modification of animal and human cells having SEQ ID No. 1, and methods for its synthesis, involving constructing vectors containing a promoter region of human elongation factor EF1A, a polylinker with sites for restriction endonucleases, a transcription terminator, a polyadenylation sequence of human growth factor, a regulatory element of transposon Tn10 allowing for antibiotic-free positive selection, an origin of replication, and a kanamycin resistance gene. Escherichia coli strain SCS110-AF is also provided by the present invention. The method for creating the strain involves constructing a linear DNA fragment containing regulatory element of transposon Tn10, a levansucrase gene, sacB, a chloramphenicol resistance gene, and two homologous sequences. The E. coli cells are transformed by electroporation and clones surviving chloramphenicol are chosen. The invention further discloses Escherichia coli strain SCS110-AF/VTvaf17, which carries DNA vector VTvaf17, and methods for its synthesis.