Abstract：
:Fragments of EcR and USP were cloned from two insect cell lines, Sf21 and High Five cells (derived respectively from Spodoptera frugiperda and Trichoplusia ni), using a PCR-based approach employing degenerate primers designed on the basis of conserved regions of nuclear receptors, together with 5'- and 3'-RACE. An additional orphan nuclear receptor, HR4 fragment, was cloned from High Five cells. Comparison of these fragments with Manduca sexta counterparts showed that the cloned SfEcR [ecdysone receptor (EcR) from Sf21 cells] had high similarity to MsEcR-B1, whereas the cloned SfUSP [ultraspiracle (USP) from Sf21 cells] and TnUSP (USP from High Five cells) matched more closely to MsUSP-2 than to MsUSP-1. The TnHR4 showed most similarity to a recently cloned Bombyx mori GRF. While EcR and USP were constitutively expressed in both cell lines, HR4 was barely detectable by Northern blot analysis in High Five cells. Treatment with 20-hydroxyecdysone (20E) and agonist RH-5992 enhanced transcription of EcR in both cell lines, while the transcription of USP was suppressed in High Five cells. Such suppressed USP transcription was not observed in Sf21 cells. Transcription of TnEcR could also be enhanced by ecdysone and 3-dehydroecdysone, whereas transcription of SfEcR was unchanged with these two ecdysteroid compounds. Induction of HR4 transcription was also observed with 20E, RH-5992, ecdysone and 3-dehydroecdysone. The protein synthesis inhibitor, cycloheximide, superinduced expression of EcR and HR4 and restored the 20E/RH-5992-suppressed expression of TnUSP in the cells. Northern blot analysis also revealed that PCR, using degenerate USP primers, was able to amplify some other orphan nuclear receptors and their expression was inducible by 20E and RH-5992 and some of them were superinducible by cycloheximide.
journal_name
Insect Biochem Mol Bioljournal_title
Insect biochemistry and molecular biologyauthors
Chen JH,Turner PC,Rees HHdoi
10.1016/s0965-1748(01)00144-8subject
Has Abstractpub_date
2002-06-01 00:00:00pages
657-67issue
6eissn
0965-1748issn
1879-0240pii
S0965174801001448journal_volume
32pub_type
杂志文章abstract：:Orcokinin (NFDEIDRSGFGFN) and orcokinin homologues are crustacean peptides eliciting potent myotropic effects in gut tissues. Through HPLC purification of brain extract of the cockroach Blattella germanica, we isolated the first insect orcokinin (NFDEIDRSGFNS). This insect orcokinin-like peptide do not show myotropic ...
journal_title：Insect biochemistry and molecular biology
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journal_title：Insect biochemistry and molecular biology
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journal_title：Insect biochemistry and molecular biology
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journal_title：Insect biochemistry and molecular biology
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journal_title：Insect biochemistry and molecular biology
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journal_title：Insect biochemistry and molecular biology
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更新日期：2006-07-01 00:00:00
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journal_title：Insect biochemistry and molecular biology
pub_type： 杂志文章
doi：10.1016/s0965-1748(99)00120-4
更新日期：2000-03-01 00:00:00
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pub_type： 杂志文章
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更新日期：1993-06-01 00:00:00
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journal_title：Insect biochemistry and molecular biology
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journal_title：Insect biochemistry and molecular biology
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更新日期：2017-12-01 00:00:00
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journal_title：Insect biochemistry and molecular biology
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pub_type： 杂志文章
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pub_type： 杂志文章
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pub_type： 杂志文章
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更新日期：2009-12-01 00:00:00
abstract：:In the Apis mellifera post-genomic era, RNAi protocols have been used in functional approaches. However, sample manipulation and invasive methods such as injection of double-stranded RNA (dsRNA) can compromise physiology and survival. To circumvent these problems, we developed a non-invasive method for honeybee gene k...
journal_title：Insect biochemistry and molecular biology
pub_type： 杂志文章
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更新日期：2009-02-01 00:00:00
