San Diego State University Research Foundation

Elitra Pharmaceuticals Inc.

Judith W Zyskind

R Allyn Forsyth

Kari Lynn Ohlsen

John D Trawick

Jamie M Froelich

Robert T Yamamoto

H Howard Xu

Grant J Carr

A method for identifying endogenous microbial proliferation genes for growth and viability is disclosed herein. The method involves exogenous nucleic acids that are used to conditionally produce antisense inhibitors of endogenous complementary mRNAs in a microorganism. Antisense fragments that result in lethality when expressed indicate that the endogenous gene is a proliferation gene. The method can also be used with sequences in sense orientation. The strategy can be used to identify new gene targets for novel antibiotics.

Methods for identifying anti-microbial agents

The present invention relates to reporter gene constructs encoding a cytoplasmic form of chitobiase (N,N'-diacetylchitobiase) and methods of using these reporter gene constructs. The use of a cytoplasmic form of chitobiase as a reporter enzyme is generally applicable to the study of gene expression in organisms which do not contain N-acetyl-&#x3b2;-D-glucosaminidases.

Chitobiase as a reporter enzyme

The sequences of nucleic acids encoding proteins required for proliferation are disclosed. The nucleic acids can be used to express proteins or portions thereof, to obtain antibodies capable of specifically binding to the expressed proteins, and to use those expressed proteins as a screen to isolate candidate molecules for rational drug discovery programs. The nucleic acids can also be used to screen for homologous genes that are required for proliferation in microorganisms other than The nucleic acids can also be used to design expression vectors and secretion vectors. The nucleic acids of the present invention can also be used in various assay systems to screen for proliferation required genes in other organisms as well as to screen for antimicrobial agents.

Genes identified as required for proliferation in Escherichia coli

The present invention relates to methods of measuring cellular proliferation using ectoenzymes such as membrane-bound chitobiase (N,N&prime;-diacetylchitobiase) and nucleic acids for use in such methods.

Use of ectoenzymes and secreted enzymes to monitor cellular proliferation

The sequences of nucleic acids encoding proteins required for proliferation are disclosed. The nucleic acids can be used to express proteins or portions thereof, to obtain antibodies capable of specifically binding to the expressed proteins, and to use those expressed proteins as a screen to isolate candidate molecules for rational drug discovery programs. The nucleic acids can also be used to screen for homologous genes that are required for proliferation in microorganisms other than . The nucleic acids can also be used to design expression vectors and secretion vectors. The nucleic acids of the present invention can also be used in various assay systems to screen for proliferation required genes in other organisms as well as to screen for antimicrobial agents.

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La Jolla, CA, United States of America

Judith W Zyskind