The United States of America As Represented by the Department of Health

Centers for Disease Control and Prevention

Frederick D Quinn

Kristin A Birkness

Michael G Schmidt

C Harold King

Marie U Owens

An isolated nucleic acid encoding a SecA protein of M. tuberculosis is provided. This nucleic acid can be a native coding sequence for the SecA protein of M. tuberculosis. A specific example of the isolated nucleic acid is one that encodes a polypeptide comprising the amino acid sequence set forth in SEQ ID NO:2. An isolated fragment of the secA gene that is specific for M. tuberculosis is provided. A purified SecA protein of Mycobacterium tuberculosis is provided. The purified SecA protein of Mycobacterium tuberculosis comprises the polypeptide having the sequence set forth in the Sequence Listing as SEQ ID NO:2. Fragments of the M. tuberculosis SecA protein are provided. A purified mutant SecA protein of Mycobacterium tuberculosis is provided. The invention provides purified mutant M. tuberculosis expressing the mutant SecA protein of the invention. The invention also provides methods of screening for putative M. tuberculosis virulence factors translocated by the SecA protein.

SecA gene of mycobacterium tuberculosis and related methods and

The present invention provides an artificial organ system comprising an endothelial cell layer, an epithelial cell layer and an artificial microporous membrane, having pores therein, disposed between and in direct contact with the endothelial cell layer and the epithelial cell layer such that the membrane has an endothelial side and an epithelial side. The present invention also provides an artificial organ system contained in a vessel comprising an upper chamber into which the epithelial side faces and containing the epithelial cell layer, and a lower chamber into which the endothelial side faces and containing the endothelial cells. The present invention also provides an artificial lung system comprising an endothelial cell layer, an alveolar epithelial cell layer and an artificial microporous membrane, having pores therein, disposed between and in direct contact with the endothelial cell layer and the alveolar epithelial cell layer such that the membrane has an endothelial side and an epithelial side. A method is also provided for constructing an artificial lung system, comprising placing an artificial microporous membrane, having pores therein, into a vessel having a bottom and supporting the membrane a distance from the bottom of the vessel to create an upper and lower chamber in the vessel such that the membrane has an endothelial side facing into the lower chamber of the vessel and an opposite epithelial side facing into the upper chamber of the vessel; placing endothelial cells into the upper chamber of the vessel under conditions such that the endothelial cells form a confluent layer of cells on the epithelial side of the membrane; and placing alveolar epithelial cells into the upper chamber of the vessel under conditions such that the endothelial cells migrate through the pores in the membrane and attach to the endothelial side of the membrane to form a confluent layer of the endothelial cells on the endothelial side of the membrane in the lower chamber and the alveolar epithelial cells form a confluent layer of the epithelial cells on the epithelial side of the membrane in the upper chamber.

Methods and compositions for an artificial lung organ culture system

The present invention provides an artificial organ system comprising an endothelial cell layer, an epithelial cell layer and an artificial microporous membrane disposed between and indirect contact with the endothelial cell layer and epithelial cell layer such that the membrane has an endothelial side and an epithelial side. Also provided is a method of constructing an artificial organ system, comprising the steps of placing an artificial microporous membrane into a tissue culture well and supporting the membrane a selected distance from a bottom of the well to create an upper and lower chamber in the well such that the membrane has an endothelial side facing the bottom of the well and an opposite epithelial side; placing endothelial cells into the upper chamber of the well under conditions such that the endothelial cells form a confluent layer of cells on the epithelial side of the membrane; and placing epithelial cells into the upper chamber of the well under conditions such that the endothelial cells migrate through pores in the membrane and attach to the endothelial side of the membrane to form a confluent layer of endothelial cells on the endothelial side of the membrane and the epithelial cells form a confluent layer of epithelial cells on the epithelial side of the membrane.

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Decatur, GA, United States of America

Frederick D Quinn