Recombinant DNA cloning vectors and the eukaryotic and prokaryot

Details Recombinant DNA cloning vectors and the eukaryotic and prokaryot Recombinant DNA cloning vectors and the eukaryotic and prokaryot

1983-09-30

1988-02-23

Martinell, James

Chemistry: molecular biology and microbiology

Spore forming or isolating process

435 68, 435 91, 4351721, 4351723, 435194, 435243, 435253, 435254, 435255, 4353171, 435320, 536 27, 935 11, 935 14, 935 29, 935 34, 935 70, 935 71, 935 72, 935 73, 935 68, 935 84, C12N 500, C12N 1500, C12N 912, C12N 100, C12N 120, C12N 114, C12N 116, C12P 2100, C12P 1934, C07H 2104

Patent

active

047270284

ABSTRACT:
The present invention comprises novel recombinant DNA cloning and expression vectors which confer hygromycin B and/or G418 resistance to eukaryotic and prokaryotic host cells. The novel recombinant DNA vectors are derived from plasmid pKC203, a plasmid which can be isolated from E. coli JR225 (ATCC 31912). The hygromycin B and G418 resistance-conferring genes can be isolated on the 7.5 kb BglII restriction fragment or the 2.5 kb SalI-BglII restriction fragment of plasmid pKC203. The eukaryotic recombinant DNA vectors of the present invention are prepared by inserting such resistance-conferring restriction fragments into a vector, such as plasmid pSV5gpt, that comprises a eukaryotic promoter and the necessary functions for maintenance of the vector as an extrachromosomal element or as an integrated sequence in the host cell chromosomal DNA. Furthermore, the present invention comprises useful derivatives of plasmid pKC203 which, although comprising no eukaryotic elements, are useful recombinant vectors for E. coli and other prokaryotes and serve as starting material for the construction of eukaryotic vectors that confer hygromycin B and/or G418 resistance to eukaryotic host cells. One useful derivative of plasmid pKC203 is constructed by circularizing the .about.7.5 kb BglIII restriction fragment of plasmid pKC203 to form plasmid pSC701, which can be further digested with HaeII or SauIIIA1 to form smaller plasmids. The present invention also comprises the novel transformants of the aforementioned recombinant DNA vectors.

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