TY - JOUR
T1 - Protocol to obtain targeted transcript sequence data from snake venom samples collected in the Colombian field
AU - Fonseca, Alejandra
AU - Renjifo-Ibáñez, Camila
AU - Renjifo, Juan Manuel
AU - Cabrera, Rodrigo
N1 - Funding Information:
This work was supported by the Fundación Cardioinfantil 2013 grant & the COLCIENCIAS grant 0558–2015 , Bogotá, Colombia.
Publisher Copyright:
© 2018 Elsevier Ltd
PY - 2018/6/15
Y1 - 2018/6/15
N2 - Snake venoms are a mixture of different molecules that can be used in the design of drugs for various diseases. The study of these venoms has relied on strategies that use complete venom extracted from animals in captivity or from venom glands that require the sacrifice of the animals. Colombia, a country with political and geographical conflicts has difficult access to certain regions. A strategy that can prevent the sacrifice of animals and could allow the study of samples collected in the field is necessary. We report the use of lyophilized venom from Crotalus durissus cumanensis as a model to test, for the first time, a protocol for the amplification of complete toxins from Colombian venom samples collected in the field. In this protocol, primers were designed from conserved region from Crotalus sp. mRNA and EST regions to maximize the likelihood of coding sequence amplification. We obtained the sequences of Metalloproteinases II, Disintegrins, Disintegrin-Like, Phospholipases A2, C-type Lectins and Serine proteinases from Crotalus durissus cumanensis and compared them to different Crotalus sp sequences available on databases obtaining concordance between the toxins amplified and those reported. Our strategy allows the use of lyophilized venom to obtain complete toxin sequences from samples collected in the field and the study of poorly characterized venoms in challenging environments.
AB - Snake venoms are a mixture of different molecules that can be used in the design of drugs for various diseases. The study of these venoms has relied on strategies that use complete venom extracted from animals in captivity or from venom glands that require the sacrifice of the animals. Colombia, a country with political and geographical conflicts has difficult access to certain regions. A strategy that can prevent the sacrifice of animals and could allow the study of samples collected in the field is necessary. We report the use of lyophilized venom from Crotalus durissus cumanensis as a model to test, for the first time, a protocol for the amplification of complete toxins from Colombian venom samples collected in the field. In this protocol, primers were designed from conserved region from Crotalus sp. mRNA and EST regions to maximize the likelihood of coding sequence amplification. We obtained the sequences of Metalloproteinases II, Disintegrins, Disintegrin-Like, Phospholipases A2, C-type Lectins and Serine proteinases from Crotalus durissus cumanensis and compared them to different Crotalus sp sequences available on databases obtaining concordance between the toxins amplified and those reported. Our strategy allows the use of lyophilized venom to obtain complete toxin sequences from samples collected in the field and the study of poorly characterized venoms in challenging environments.
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U2 - 10.1016/j.toxicon.2018.03.009
DO - 10.1016/j.toxicon.2018.03.009
M3 - Research Article
C2 - 29571682
AN - SCOPUS:85045199843
SN - 0041-0101
VL - 148
SP - 1
EP - 6
JO - Toxicon
JF - Toxicon
ER -