Int J Biol Sci 2009; 5(5):451-457. doi:10.7150/ijbs.5.451
Short Research Communication
Construction of a full-length cDNA Library from Chinese oak silkworm pupa and identification of a KK-42-binding protein gene in relation to pupa-diapause termination
1. College of Bioscience and Biotechnology, Shenyang Agricultural University, Shenyang 110161, China
In this study we successfully constructed a full-length cDNA library from Chinese oak silkworm, Antheraea pernyi, the most well-known wild silkworm used for silk production and insect food. Total RNA was extracted from a single fresh female pupa at the diapause stage. The titer of the library was 5 × 105 cfu/ml and the proportion of recombinant clones was approximately 95%. Expressed sequence tag (EST) analysis was used to characterize the library. A total of 175 clustered ESTs consisting of 24 contigs and 151 singlets were generated from 250 effective sequences. Of the 175 unigenes, 97 (55.4%) were known genes but only five from A. pernyi, 37 (21.2%) were known ESTs without function annotation, and 41 (23.4%) were novel ESTs. By EST sequencing, a gene coding KK-42-binding protein in A. pernyi (named as ApKK42-BP; GenBank accession no. FJ744151) was identified and characterized. Protein sequence analysis showed that ApKK42-BP was not a membrane protein but an extracellular protein with a signal peptide at position 1-18, and contained two putative conserved domains, abhydro_lipase and abhydrolase_1, suggesting it may be a member of lipase superfamily. Expression analysis based on number of ESTs showed that ApKK42-BP was an abundant gene in the period of diapause stage, suggesting it may also be involved in pupa-diapause termination.
Keywords: Chinese oak silkworm, Antheraea pernyi, cDNA library, Expressed sequence tag, KK-42-binding protein, diapause termination
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How to cite this article:
Li YP, Xia RX, Wang H, Li XS, Liu YQ, Wei ZJ, Lu C, Xiang ZH. Construction of a full-length cDNA Library from Chinese oak silkworm pupa and identification of a KK-42-binding protein gene in relation to pupa-diapause termination. Int J Biol Sci 2009; 5(5):451-457. doi:10.7150/ijbs.5.451. Available from http://www.ijbs.com/v05p0451.htm