Screening of silkworm strains for efficient recombinant protein production by Autographa californica nucleopolyhedrosis virus (AcNPV)
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Baculoviruses base vectors come to be regarded as methods for in vivo gene deliveryand transient expression to the silkworm. In the case of silkworm, B. mori, two types ofbaculoviruses, AcNPV (Autographa californica nuclear polyhedrosis virus) and BmNPV(Bombyx mori nuclear polyhedrosis virus), are potentially applicable as vectors. Recently,AcNPV showed promising results with some silkworm strains despite different hostspecificities. We searched for a highly-permissive silkworm strain in the B. mori stocksof Kyungpook National University that could produce high levels of recombinant protein. Seventy strains were screened using the recombinant AcNPV/BmA3-Luc virus. Based on themeasured luciferase activity, the strains could be divided into three groups, high-, middle-, andlow-permissive strains, according to their relative recombinant protein expression levels. At 48hours post-injection, the luciferase activity in the high-permissive strains was 500-fold greaterthan that of the low-permissive strains. At 72 hours post-injection, a significant elevation inluciferase activity was observed in the hemocytes of all strains. Then, based on the aboveresults, the High Permissive Strain (HPS) S10 and the Low Permissive Strain (LPS) S39 werepick up and was carried out Dot blotting, RT-PCR and Real time PCR.
Abstract
Baculoviruses base vectors come to be regarded as methods for in vivo gene deliveryand transient expression to the silkworm. In the case of silkworm, B. mori, two types ofbaculoviruses, AcNPV (Autographa californica nuclear polyhedrosis virus) and BmNPV(Bombyx mori nuclear polyhedrosis virus), are potentially applicable as vectors. Recently,AcNPV showed promising results with some silkworm strains despite different hostspecificities. We searched for a highly-permissive silkworm strain in the B. mori stocksof Kyungpook National University that could produce high levels of recombinant protein. Seventy strains were screened using the recombinant AcNPV/BmA3-Luc virus. Based on themeasured luciferase activity, the strains could be divided into three groups, high-, middle-, andlow-permissive strains, according to their relative recombinant protein expression levels. At 48hours post-injection, the luciferase activity in the high-permissive strains was 500-fold greaterthan that of the low-permissive strains. At 72 hours post-injection, a significant elevation inluciferase activity was observed in the hemocytes of all strains. Then, based on the aboveresults, the High Permissive Strain (HPS) S10 and the Low Permissive Strain (LPS) S39 werepick up and was carried out Dot blotting, RT-PCR and Real time PCR.
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