SBIR-STTR Award

The objective of this project is to construct a mycomplasma gallisepticum biotinylated DNA probe. Economic losses from m. Gallisepticum infection are estimated to be $118 million annually in the United States. Minimum efficiency of M. Gallisepticum disease control requires a rapid and sensitive identification system such as a probe. Mycoplasma Gallisepticum DNA will be labeled will biotin-11-dutp in a standard nick-translation reaction. Target DNA hybridize ) with the biotinylated probe on a nitrocellulose filter will be incubated with streptavidin and polyalkaline phosphatase. Upon addition of an enzyme/substrate complex, the hybridized DNA will appear as a blue spot. Results can be obtained in 24-36 hours, which is considerably faster than routine methods currently in use.

Keywords:
1. Biotechnology and genetic engineering

The object of this project is to construct a specific MvcoDlasma RalliseDticum biotinylated DNA probe. Economic losses from M. YalliseDticum infection are estimated to be $118 million annually in the United States. Maximum efficiency of M. ralliseoticum disease control requires a rapid and sensitive identification system such as a probe. Total M. œalliseDticum genome DNA was labeled with biotin-ll-dUTP. It is capable of detecting 75 ng of DNA or 7.5 x 103 broth-grown organisms by hybridization reaction in 19 hours. This probe also detects many other mycoplasmas. MvooDlasma RalliseDticum species-specificity will be accomplished by cloning an M RalliseDticum-specific DNA sequence into Charon 4A and using this as the DNA source for the biotinylated probe.