Unmasking venom gland transcriptomes in reptile venoms.
While structural studies of reptile venom toxins can be achieved using lyophilized venom samples, until now the cloning of precursor cDNAs required sacrifice of the specimen for dissection of the venom glands. Here we describe a simple and rapid technique that unmasks venom protein mRNAs present in lyophilized venom samples. To illustrate the technique we have RT-PCR-amplified a range of venom protein transcripts from cDNA libraries derived from the venoms of a hemotoxic snake, the Chinese copperhead (Deinagkistrodon acutus), a neurotoxic snake, the black mamba (Dendroaspis polylepis), and a venomous lizard, the Gila monster (Heloderma suspectum). These include a metalloproteinase and phospholipase A2 from D. acutus, a potassium channel blocker, dendrotoxin K, from D. polylepis, and exendin-4 from H. suspectum. These findings imply that the apparent absence and/or lability of mRNA in complex biological matrices is not always real and paves the way for accelerated acquisition of molecular genetic data on venom toxins for scientific and potential therapeutic purposes without sacrifice of endangered herpetofauna.
Chen
Tianbao
T
Pharmaceutical Biotechnology, School of Biomedical Sciences, University of Ulster, Coleraine BT52 1SA, Ireland.
Bjourson
Anthony J
AJ
Orr
David F
DF
Kwok
HangFai
H
Rao
Pingfan
P
Ivanyi
Craig
C
Shaw
Chris
C
D000818:Animals; D003001:Cloning, Molecular / Q000379:methods; D018076:DNA, Complementary / Q000096:biosynthesis / Q000302:isolation & purification*; D000077270:Exenatide; D005612:Freeze Drying; D008116:Lizards; D008666:Metalloendopeptidases / Q000235:genetics; D010455:Peptides / Q000235:genetics; D010741:Phospholipases A / Q000235:genetics; D054467:Phospholipases A2; D012333:RNA, Messenger / Q000302:isolation & purification; D012910:Snake Venoms / Q000235:genetics; D014688:Venoms / Q000235:genetics*
D018076:DNA, Complementary; D010455:Peptides; D012333:RNA, Messenger; D012910:Snake Venoms; D014688:Venoms; C081506:dendrotoxin K; D000077270:Exenatide; D010741:Phospholipases A; D054467:Phospholipases A2; D008666:Metalloendopeptidases