Purified bacterial protein which stimulates ligation of nucleic acid molecules

• US 5,229,273 A
• Filed: 05/30/1990
• Issued: 07/20/1993
• Est. Priority Date: 05/30/1990
• Status: Expired due to Fees

• Current Assignee: Trustees Of Columbia University In The City Of New York (Columbia University)
• Original Assignee: Trustees Of Columbia University In The City Of New York (Columbia University)
• Inventors: Holloman, William, Gottesman, Max
• Primary Examiner(s): Wax, Robert A.
• Assistant Examiner(s): WALSH, STEPHEN G
• Application Number:
  US07/530,922
• Time in Patent Office: 1,147 Days
• Field of Search: 530/350, 530/825, 536/27, 435/69.1, 435/71.2, 435/320.1, 435/252.33, 435/183
• US Class Current: 435/69.1
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C12N 15/70 Vectors or expression systems specially adapted for E. coli
• CPC Class Codes: C12N 2795/10122 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes

Abstract

The subject invention provides a purified protein having a molecular weight of about 22.8 kDa and the amino acid sequence shown in FIG. 2. The invention further provides a method of producing the 22.8 kDa protein which comprises culturing the host cell comprising an expression vector under conditions such that the expression vector contained therein expresses the protein and recovering from the host cell the protein so expressed. Also provided is a method of ligating separate nucleic acid molecules which comprises contacting the molecules with the 22.8 kDa protein under conditions such that the molecules are ligated together.

Claims

1. An expression plasmid designated pLSD13.

2. An Escherichia coli 7298 cell (ATCC Accession No. 68326) comprising the pLSD13 expression plasmid of claim 1.

3. A method of producing a purified protein having a molecular weight of about 22.8 kD, the amino acid sequence shown in FIG. 2 and the ability to ligate double-stranded DNA molecules, which comprises culturing the Escherichia coli 7298 cell of claim 2 under conditions such that the pLSD13 plasmid contained therein expresses the protein and then recovering the protein so expressed.