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Molecular cloning and expression in vitro of a carboxylesterase gene from the Glanville fritillary butterfly (Melitaea cinxia)
Authors:Shiqi Luo  Chang ShuChongren Xu  Rongjiang Wang
Affiliation:College of Life Sciences, Peking University, Beijing 100871, PR China
Abstract:Carboxylesterase (EC 3.1.1.1) is a member of the carboxyl/cholinesterase (CCE) superfamily, which is widely distributed in animals, plants and microorganisms. This enzyme has been known to be associated with insecticide resistance and detoxification. Although CCEs have been extensively studied in insects, including lepidopterans, the research on butterflies, a major subgroup in Lepidoptera, is still poor. In the present study, we cloned a CCE gene (McCCE1) from the Glanville fritillary butterfly (Melitaea cinxia, Lepidoptera: Nymphalidae). The full-length cDNA encoding McCCE1 was 1786 bp, containing a 1641 bp open reading frame encoding 546 amino acids, a 38 bp 5′-untranslated region (5′-UTR), and a 107 bp 3′-UTR with a poly(A) tail. The functionally conserved amino acids in McCCE1 shared the 55% identity with the cytoplasmic esterase CCE017a in Helicoverpa armigera (Lepidoptera: Noctuidae), which has been associated with detoxification. Assays in vitro showed that the recombinant McCCE1 could hydrolyze α- and β-naphthyl acetate. Thus, the present study adds to the body of knowledge concerning the detoxification of pesticides by lepidopterans.
Keywords:bp, base pairs   CCE, carboxyl/cholinesterase   cDNA, complementary DNA   CXE3, carboxylesterase 3   dNTP, deoxyribonucleoside triphosphate   kDa, kilodalton(s)   ORF, open reading frame   PAGE, polyacrylamide gel electrophoresis   PCR, polymerase chain reaction   RACE, rapid amplification of cDNA ends   UTR, untranslated region
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