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Marine Ecology Progress Series

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MEPS 376:123-132 (2009)  -  DOI:

Quantification of coral heat shock proteins from individual coral polyps

Erin Bromage1,3,*,**, Lawrence Carpenter2,4,**, Stephen Kaattari1, Mark Patterson2

1Department of Environmental and Aquatic Animal Health, and 2Department of Biological Sciences, Virginia Institute of  Marine Science, College of William & Mary, Gloucester Point, Virginia 23062, USA
3Department of Biology, University of Massachusetts Dartmouth, 285 Old Westport Road, North Dartmouth, Massachusetts 02747, USA
4Applied Ocean Physics & Engineering Department, Woods Hole Oceanographic Institution, Deep Submergence Laboratory,  MS#7, 266 Woods Hole Road, Woods Hole, Massachusetts 02543, USA
**These authors contributed equally to this work

ABSTRACT: The induction and regulation of heat shock proteins (hsps) is a significant defense mechanism that can preserve metabolic function and foster recovery from short-term stress events. Present coral sampling methodologies that involve hsp analysis often require the harvesting of large samples of live coral colonies that may already be stressed or in poor health. In the present study, 3 novel protocols were developed to: (1) extract single coral polyps, minimizing colony trauma; (2) purify protein from single coral polyps (approximately 12 mm3); and (3) develop a more sensitive protein quantification method. The preliminary testing of 5 separate protein preparation methods resulted in a range of total protein yields from 47 to 77 µg coral polyp–1. The optimized methods were able to recover, on average, 44 ± 12 µg protein polyp–1 (n = 20). Subsequent SDS-PAGE and immunoblotting analysis of single coral polyps resolved as little as 87 pg hsp70 coral polyp–1. This minimally invasive sampling protocol reduces coral damage and, thus, reduces stress and diseases caused by sampling.

KEY WORDS: Coral · Heat stock proteins · hsp · Polyp · Proteins · Stress

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Cite this article as: Bromage E, Carpenter L, Kaattari S, Patterson M (2009) Quantification of coral heat shock proteins from individual coral polyps. Mar Ecol Prog Ser 376:123-132.

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