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

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...

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Bibliographic Details
Published in:International journal of biological sciences 2009-01, Vol.5 (5), p.451-457
Main Authors: Li, Yu-Ping, Xia, Run-Xi, Wang, Huan, Li, Xi-Sheng, Liu, Yan-Qun, Wei, Zhao-Jun, Lu, Cheng, Xiang, Zhong-Huai
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Antheraea pernyi
Base Sequence
Bombyx mori
Expressed Sequence Tags
Gene Library
Genes, Insect
Imidazoles - metabolism
Insect Proteins - genetics
Insect Proteins - metabolism
Juvenile Hormones - metabolism
Metamorphosis, Biological - genetics
Molecular Sequence Data
Moths - genetics
Moths - metabolism
Pupa
Short Research Communication
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Summary: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 x 10(5) 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.
ISSN:1449-2288
1449-2288
DOI:10.7150/ijbs.5.451