Protein fusion method and constructs

Details Protein fusion method and constructs Protein fusion method and constructs

1995-01-25

1999-11-09

Ketter, James

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

4351723, 43525233, 4353201, 536 231, C12Q 168, C12N 121, C12N 1500

Patent

active

059811773

ABSTRACT:
The instant disclosure teaches transposons and methods of the instant invention for making Protein Fusions by rapid, random shuffling of protein domains to produce novel protein fusions. This system is generally applicable to production of multifunctional chimeric proteins in vivo and in vitro. The methods and constructs of the instant invention can be used to randomly create both carboxy- and amino-terminal protein fusions in vivo. The methods and contructs of the instant invention are useful in the development of a protein domain library, in the construction of multifunctional enzymes, and in the accelerated evolution of new enzymatic activities.

REFERENCES:
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Manoil, Analysis of Protein Localization by Use of Gene Fusions with Complementary Properties, J. Bacteriol. 172(2): 1035-1042, Feb. 1990.
Clark et al., Transcription of the Escherichia coli recE gene from a promoter in Tn5 and IS50, J. Biol. Chem. 176(22): 7024-7031, Oct. 1994.
Camilli et al., Insertional Mutagenesis of Listeria monocytogenes with a novel Tn917 Derivative that allows direct cloning of DNA flanking transposon insertions, J. Bacteriol. 172(7): 3738-3744, Jul. 1990.
Bhagwat et al., Prinary sequence of the EcoRII endonuclease and properties of its fusions with Beta-galactosidase, J. Biol. Chem. 265(2): 767-773, Jan. 1990.
Phage Mu edited by N. Symonds, A. Toussaint, P. van de Putte, and M. Howe (Cold Spring Harbor Laboratory, 1987). Chapter 2.
Phage Mu edited by N. Symonds, A. Toussaint, P. van de Putte, and M. Howe (Cold Spring Harbor Laboratory, 1987). Chapter 12.
Phage Mu edited by N. Symonds, A. Toussaint, P. van de Putte, and M. Howe (Cold Spring Harbor Laboratory, 1987). Chapter 14.

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