Jan Kostal

US Patent:

20060040393, Feb 23, 2006

Filed:

Aug 16, 2005

Appl. No.:

11/205867

Inventors:

Xiyu Jia - Newport Beach CA, US
Jan kostal - Saint Paul MN, US
Jonathan Claypool - Seattle WA, US

International Classification:

C12N 15/74
C12N 1/21

US Classification:

435488000, 435252330

Abstract:

The present invention relates to the simple, gentle, and efficient extraction of biological material from (). The use of in research laboratories depends on the ability to prepare lysates to isolate the desired products under investigation. The present invention includes methods and engineered strains that are capable of rapid controlled lysis or herein “autolysis”. The XJa strains were made from JM109 and the XJb strains from BL21 by insertion of the λ R or (λ RS) lytic endolysin gene to replace the tightly regulated araB gene. Thus, arabinose becomes a non-metabolizable inducer and the controlled autolysis phenotype is induced by the Ppromoter by the presence of saturating arabinose. Upon induction of the bacteriophage λR endolysin, the remains intact but is efficiently lysed after one freeze-thaw cycle. The present invention is usable with many different buffer systems and is flexible in this regard. The controlled autolysis phenotype shows increrased yields and purity of extracted protein compared to detergent based lysis or traditional sonication lysis methods. The present invention is useful for routine protein expression or nucleic acid extraction and also for high-through-put manipulation involving protein or nucleic acid from