AU 555574 B

Actual granted independent claims

AU 555574 B

Claim 1
A recombinant DNA plasmid which comprises:
a) a eukaryotic promoter;
b) one or two different structural genes and associated control sequence that convey resistance to either or both of antibiotics hygromycin B and G418 when transformed into a host cell that is sensitive to either or both antibiotics for which resistance is conveyed, said host cell being susceptible to transformation, cell division, and culture; and
c) a prokaryotic replicon, said replicon being functional when said host cell is prokaryotic, subject to the limitations

  • that the one or two structural genes and associated control sequence are adjacent to and, in a eukaryotic host cell, transcribed from the eukaryotic promoter,
  • that a single gene and associated control sequence conveys resistance to either but not both hygromycin B and G418, and
  • that the gene conveying resistance to G418 does not code for the enzyme phosphotransferase.
Claim 19A recombinant DNA cloning vector substantially as described with reference to Examples 3, 7, 10, 16, 19, 22, 30, 33, 35, 37, 39, and 4112.
12 The examples refer to:
Ex. 3, 7 and 10: construction of plasmids pKC214, pKC215, pGD1, pGD2, pGD3 and pGD4 with the 7.5 kb Blg II fragment having both hygromycin and G418 resistance genes in different orientations.
Ex. 16: construction of plasmids pGD10 and pGD11 with the 1.51 kb Sac I/Blg II fragment having a hygromycin resistance gene in different orientations.
Ex. 19: construction of plasmids pGD12 and pGD13 with the 1.65 kb EcoR I/Sal I fragment having a G418 resistance gene in different orientations.
Ex. 22: construction of plasmids pGD14 and pGD15 with the 2.5 kb Sal I/Bgl II fragment having a G418 resistance gene in different orientations.
Ex. 30: construction of plasmids pLO314 and pLO315 with both hygromycin and G418 resistance genes in different orientations functional in prokaryotes and eukaryotes.
Ex. 33, 35, and 37: construction of plasmids pLO316, pLO317, pLO318, pLO319, pLO320 and pLO321 with hygromycin resistance gene in different orientations functional in prokaryotes and eukaryotes.
Ex. 39 and 41: construction of plasmids pKC273 and pLO378 with both hygromycin and G418 resistance genes and functional in prokaryotes and eukaryotes.
Claim 20A transformed host cell substantially as described with reference to Examples 3 to 5, 7 to 12, 16 to 24, and 31 to 4213 . 13 The transformed host cells referred to in the examples are:
Ex. 3 to 5: Mouse cells.
Ex. 7 to 12: Mouse, yeast and E. coli cells.
Ex. 16 to 24: Mouse and E. coli cells.
Ex. 31 to 42: Mouse, Saccharomyces and E. coli