期刊
EUROPEAN JOURNAL OF ENTOMOLOGY
卷 108, 期 1, 页码 164-168出版社
CZECH ACAD SCI, INST ENTOMOLOGY
DOI: 10.14411/eje.2011.020
关键词
Universal primer; Lepidoptera; coding sequence; untranslated region; HSP90; RACE
类别
资金
- National Natural Science Foundation of China (NSFC) [30770302, 30570970]
- Ministry of Science and Technology of the Republic of China [2006FY110500]
- National Science Fund for Fostering Talents in Basic Research [NSFC-J0630964/J0109]
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.
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