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Issue:ISSN 2095-1353
           CN 11-6020/Q
Director:Chinese Academy of Sciences
Sponsored by:Chinese Society of Entomological;institute of zoology, chinese academy of sciences;
Address:Chaoyang District No. 1 Beichen West Road, No. 5 hospital,Beijing City,100101, China
Your Position :Home->Past Journals Catalog->2014年51 No.4

Molecular cloning and expression profiles of a Kazal-type serine protease inhibitor gene in Antheraea pernyi
Author of the article:WANG Lei** QIU Jian-Feng QIAN Cen ZHU Bao-Jian LIU Chao-Liang
Author's Workplace:College of life science, Anhui Agricultural University, Hefei 230036, China
Key Words:Antheraea pernyi, Kazal, gene cloning, gene expression
Abstract:[Objectives]  To clone full-length cDNA of a Kazal-type serine proteinase inhibitor in Antheraea pernyi (ApKTSPI), to analyze its expression level in tissue and in fat bodies after a pathogen’s immune-challenge, and to get it expressed in Escherichia coli. [Methods]  ApKTSPI cDNA was amplified using the RACE-PCR method and the expression pattern analyzed using quantitative real time PCR. The ApKTSPI gene was inserted into the pET-28a vector. Recombinant plasmids were transfected into E. coli BL21, and the ApKTSPI protein was expressed with IPTG induction. [Results]  The full-length cDNA of ApKTSPI was 568 nucleotides long and contained a 291 nucleotide ORF encoding a 96-residue amino acid sequence. The ApKTSPI gene was especially high expressed in fat bodies of the 5th larval instar. The ApKTSPI gene in fat bodies was up-regulated after challenging the immune system with NPV, E. coli or Beauveria bassiana. However, the degree and timing of up-regulation were different for each of the three pathogens. The recombinant ApKTSPI protein was successfully expressed in E. coli. [Conclusion]  The ApKTSPI gene was successfully cloned and expressed in E. coli, and ApKTSPI may be involved in innate immune reactions against pathogens.
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