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Characterization and functional analysis of serpin-1 like gene from oak silkworm Antheraea pernyi

Published online by Cambridge University Press:  23 February 2017

H.M. Yu ,
B.J. Zhu ,
Y. Sun ,
G.Q. Wei ,
L. Wang ,
C. Qian ,
M. Nadeem Abbas  and
C.L. Liu
H.M. Yu
Affiliation:
College of Life Sciences, Anhui Agricultural University, Hefei, 230036, China
B.J. Zhu
Affiliation:
College of Life Sciences, Anhui Agricultural University, Hefei, 230036, China
Y. Sun
Affiliation:
College of Life Sciences, Anhui Agricultural University, Hefei, 230036, China
G.Q. Wei
Affiliation:
College of Life Sciences, Anhui Agricultural University, Hefei, 230036, China
L. Wang
Affiliation:
College of Life Sciences, Anhui Agricultural University, Hefei, 230036, China
C. Qian
Affiliation:
College of Life Sciences, Anhui Agricultural University, Hefei, 230036, China
M. Nadeem Abbas
Affiliation:
College of Life Sciences, Anhui Agricultural University, Hefei, 230036, China
C.L. Liu*
Affiliation:
College of Life Sciences, Anhui Agricultural University, Hefei, 230036, China
*
*Author for correspondence Phone: +86 551 6578 -6325 Fax: +86 551 6578-6325 E-mail: clliu@ahau.edu.cn
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Abstract

Serpins are a broadly distributed family of proteases found in various organisms that play an important role in regulating the immune response. Here, we identified a serpin-1 gene from Antheraea pernyi that encodes a 279 amino acid protein with a molecular weight of 30.8 kDa. We expressed the recombinant Ap-serpin-1 protein in Escherichia coli and used the purified protein to prepare rabbit anti-Ap-serpin-1 polyclonal antibodies. We calculated the enzyme-linked immunosorbent assay titer of the antibody as 1:128000. Quantitative real-time polymerase chain reaction analysis revealed that Ap-serpin-1 was expressed in all examined tissues, including hemolymph, malpighian tubules, midgut, silk gland, integument and the fat body; the highest Ap-serpin-1 expression levels was detected in the fat body. We next investigated the expression patterns of Ap-serpin-1 in both fat body and hemolymph samples, following treatment with E. coli, Beauveria bassiana, Micrococcus luteus and nuclear polyhedrosis virus (NPV). We reported that NPV and M. luteus significantly enhanced Ap-serpin-1 expression in the fat body. While, in the hemolymph samples, treatment with B. bassiana and M. luteus was shown to upregulate Ap-serpin-1 expression at 24 h induction. Altogether, our results suggest that Ap-serpin-1 is involved in the innate immunity of A. pernyi.

Keywords

Antheraea pernyiserpin-1innate immunityexpression
Type
Research Papers
Information
Bulletin of Entomological Research , Volume 107 , Issue 5 , October 2017 , pp. 620 - 626
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Copyright
Copyright © Cambridge University Press 2017 

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Footnotes

†

These authors contributed equally to this work.

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