Leslie M Hoffman, Age 741121 Erin St, Madison, WI 53715

7101713, Sep 5, 2006

May 16, 2002

10/147564

Leslie M. Hoffman - Madison WI, US
Jerome J. Jendrisak - Madison WI, US

Epicentre Technologies Corporation - Madison WI

C12N 15/64

435473, 435474, 435476

It can be difficult to achieve efficient transformation of many strains of bacterial cells due in part to the presence of one or more restriction and modification (R-M) systems in the cells that restricts unmodified transforming DNA. Phage T7 OCR protein is a potent inhibitor of Type I R-M systems. Methods are disclosed for improving transformation efficiency of Eubacterial and Archaebacterial cells having an R-M system by introducing into the cells an inhibitor of the restriction activity. For example, addition of 1–5 micrograms of T7 OCR protein to 50 microliters of electrocompetent cells having a Type I R-M system prior to electroporation significantly increased transformation efficiency by unmodified plasmids, fosmid clones, and artificial transposons comprising synaptic complexes. Other inhibitors or restriction activity, such as phage-encoded proteins and proteins encoded by conjugative plasmids, as well as other disclosed inhibitors, also can be used to improve transformation (and transposition) efficiency. Host cells with heritable improved transformation efficiency can be made by transforming a cell having an R-M system with an expressible gene which encodes an inhibitor of the restriction activity, which gene can be conditionally expressible.

7138267, Nov 21, 2006

Apr 4, 2002

10/116886

Jerome J. Jendrisak - Madison WI, US
Leslie M. Hoffman - Madison WI, US
Michael J. Fiandt - Cambridge WI, US
Darin Haskins - Madison WI, US

Epicentre Technologies Corporation - Madison WI

C12N 1/21
C12N 15/63

4352523, 435471

A method for retrofitting DNA in a single-copy or high-copy vector, such as a fosmid or BAC, whereby an artificial transposon is used to introduce a conditional multi-copy origin of replication (“ori”) into the DNA in said vector. Following random in vitro or in vivo transposition of the ori-containing transposon into DNA in the single-copy or low-copy vector, the resulting insertion clones are introduced into a special host strain that contains a gene which encodes a polypeptide required for replication from the multi-copy ori. However, since the gene for this polypeptide is expressed from a tightly-regulated inducible promoter, the polypeptide is not expressed in the absence of inducer. On addition of inducer to the culture medium, the host cell synthesizes the polypeptide, which in turn activates replication from the multi-copy ori, thereby increasing the amount of clone DNA synthesized by the cell.

7794971, Sep 14, 2010

Jul 1, 2004

10/883459

Darin J. Haskins - Blue Mound WI, US
Leslie M. Hoffman - Madison WI, US

Epicentre Technologies Corporation - Madison WI

C12N 15/00
C12N 15/85

435 691, 435325

The present invention provides compositions and methods for controlling the copy number for a broad range of plasmids and uses thereof. Disclosed is a host cell for conditional control of copy number of a plasmid, which host cell comprises a poly(A) polymerase gene that is operably joined to a conditionally inducible promoter, and a method for cloning and stably maintaining a DNA sequence encoding a heterologous polypeptide in the host cell.

2010032, Dec 23, 2010

Jul 15, 2010

12/837466

Darin J. Haskins - Blue Mound WI, US
Leslie M. Hoffman - Madison WI, US

C12P 21/00
C12N 1/21
C12P 19/34

435 691, 43525233, 435 9132, 435 911

The present invention provides compositions and methods for controlling the copy number for a broad range of plasmids and uses thereof. Disclosed is a host cell for conditional control of copy number of a plasmid, which host cell comprises a poly(A) polymerase gene that is operably joined to a conditionally inducible promoter, and a method for cloning and stably maintaining a DNA sequence encoding a heterologous polypeptide in the host cell.

5589616, Dec 31, 1996

Jul 25, 1994

8/279616

Leslie M. Hoffman - Madison WI

Mycogen Plant Science, Inc. - San Diego CA

C12N 514
C12N 1529
A01H 500
A01H 510

800205

The present invention discloses dicot cells containing monocot seed storage protein. Construction of genes encoding monocot seed storage proteins which are expressible in dicot cells and transformation of such genes into plant cells is also taught. Furthermore, methods and DNA molecules useful for producing plant cells containing monocot seed storage proteins are also disclosed. The invention is exemplified by combination of a 15 kD zein structural gene from a Zea mays gene with a promoter and a polyadenylation site derived from a Phaseolus vulgaris phaseolin gene.

5003045, Mar 26, 1991

Aug 29, 1986

6/902223

Leslie M. Hoffman - Madison WI

Lubrizol Genetics, Inc. - Wickliffe OH

C07K 1510

530378

The present invention discloses plant cells which contain modified 7S legume seed storage protein. Modification of 7S seed storage proteins which are expressible in plant cells and transformation of such genes into plant cells is also taught. Furthermore, methods and DNA molecules useful for producing plant cells containing modified 7S seed storage proteins are also disclosed. The invention is exemplified by insertion of an oligonucleotide encoding 15 amino acid residues, including 6 methionines, into a Phaseolus vulgaris phaseolin gene, thereby tripling its content of sulfur-containing amino acids.

6190865, Feb 20, 2001

Sep 27, 1995

8/534799

Jerome J. Jendrisak - Madison WI
Leslie M. Hoffman - Madison WI
Robert E. Smith - Madison WI

Epicentre Technologies Corporation - Madison WI

C12Q 168
C12P 1934

435 6

A method of characterizing a nucleic acid molecule is disclosed. The method comprises synthesizing DNA in the presence of a reaction mixture comprising a nucleic acid template, a primer molecule, an enzyme that extends the primer so that a DNA molecule may be synthesized, four canonical deoxynucleoside triphosphates and at least one non-canonical deoxynucleoside triphosphate. The non-canonical deoxynucleoside triphosphate is incorporated into the synthesized DNA in place of a portion of only one canonical deoxynucleoside triphosphate. The synthesized DNA is treated with an N-glycosylase that excises a base portion of the non-canonical deoxynucleoside triphosphate from the synthesized DNA. The DNA is then treated in such a manner that the phosphodiester backbone of the DNA is broken at the abasic site, thus creating at least two DNA fragments. The fragments are separated according to size.

5215912, Jun 1, 1993

Aug 14, 1989

7/393834

Leslie M. Hoffman - Madison WI

Lubrizol Genetics, Inc. - Wickliffe OH

C12N 500
C12N 1500
C12P 2100
C12P 2104

4352404

The present invention discloses dicot cells containing monocot seed storage protein. Construction of genes encoding monocot seed storage proteins which are expressible in dicot cells and transformation of such genes into plant cells is also taught. Furthermore, methods and DNA molecules useful for producing plant cells containing monocot seed storage proteins are also disclosed. The invention is exemplified by combination of a 15 kD zein structural gene from Zea mays gene with a promoter and a polyadenylation site derived from a Phaseolus vulgaris phaseolin gene.