Eur. J. Entomol. 108 (1): 164-168, 2011 | DOI: 10.14411/eje.2011.020

Universal primers for amplifying the complete coding sequence of cytoplasmic heat shock protein 90 (HSP90) in Lepidoptera

Peng Jun XU*,1,4, Tong LI*,1,4, Jin Hua XIAO1, Robert W. MURPHY3,5, Huang DA WEI**,1,2
1 Key Laboratory of Zoological Systematics and Evolution, Institute of Zoology, Chinese Academy of Sciences, 1 Beichen West Road, Chaoyang District, Beijing 100101, China
2 College of Life Sciences, Hebei University, Baoding, 071002, China
3 State Key Laboratory of Genetic Resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, China
4 Graduate School of the Chinese Academy of Sciences, Beijing, China
5 Department of Natural History, Royal Ontario Museum, 100 Queen's Park, Toronto, Ontario M5S 2C6, Canada

Using sequence alignment, a conserved domain in the 3' untranslated region (UTR) of the cytoplasmic heat shock protein 90 (HSP90) of Lepidoptera was found. This region is highly variable in other insect groups. Furthermore, universal primers were designed to amplify the complete coding sequence (CDS) of HSP90 from total genomic DNA in Lepidoptera, avoiding the commonly used reverse transcription-polymerase chain reaction (RT-PCR) and 3', 5'-rapid amplification of cDNA ends (RACE) methods based on cDNA. These primers amplified a fragment of about 2.25 kb in the 11 species tested, which represent seven different families of Lepidoptera, including moths and butterflies. The results suggest that the conserved domain of 3'UTR is universal in Lepidoptera and these primers successfully amplify the complete CDS of cytoplasmic HSP90 from genomic DNA.

Keywords: Universal primer, Lepidoptera, coding sequence, untranslated region, HSP90, RACE

Received: August 19, 2010; Accepted: October 8, 2010; Published: January 3, 2011

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